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Fulltext Evaluation of umaminess in green mussel hydrolysate (Perna viridis) produced in the presence of sodium tripolyphosphate and NaCl

Normah, I., Nurdalila Diyana, M.R.

International Food Research Journal, 2018;25(6):524-530.
MyJurnal

This study was conducted to evaluate umami taste in protein hydrolysate produced from green mussel (Perna viridis) by hydrolysing with flavorzyme at pH 8, enzyme substrate ratio (E/S) 3% with or without the presence of 0.4% sodium tripolyphosphate (STPP) and 1.5% NaCI. Degree of hydrolysis (DH), yield, amino acid compositions, molecular weight distribution and sensory evaluation were determined. The highest DH (23.18%), darkest color and highest yield (8.34%) were recorded for hydrolysate produced in the presence of both STPP and NaCI. Electrophoresis analysis showed the presence of protein bands between 10 to 70 kDa where hydrolysate with addition of STPP and NaCI had bands with lower intensities. Amino acids which contribute to the umami taste such as glutamic acid, glycine and aspartic acid were higher in hydrolysate produced with STPP and NaCI addition. The hydrolysate has lesser fishy odor and flavor than those produced with only in the presence of flavorzyme and was also rated with highest score for all the five basic tastes including bitterness. However, the score for bitterness was still lower than the reference solutions. Therefore, green mussel hydrolysate produced in this study has a good potential as a food flavorant.

Matched MeSH terms: Electrophoresis
Optimization method for proteomic analysis of the larva and adult tissues of plutella xylostella (l.) (lepidoptera: plutellidae)

Maizom Hassan, NorazilaYusoff, Wan Mohd Aizat, Idris Abd Ghani, Nurul Wahida Othman

Sains Malaysiana, 2018;47:2975-2983.

Plutella xylostella (L.) (Lepidoptera: Plutellidae), the major insect pest of cruciferous crops worldwide shows significant
resistance to almost all classes of insecticides. In order to effectively prevent and manage the insecticidal resistance,
it is crucial to understand the physiological adaptation of insects against insecticides. Identification of insect protein
that interacting with insecticides and characterization of their modification in resistant strains can be done by using
differential proteomics approach. This study focuses on optimizing a sensitive and rapid method for the extraction of
high quality protein of both larva and adult tissues of P. xylostella to be used in two-dimensional gel electrophoresis.
Five extraction methods were evaluated for protein concentration, yields and resolving patterns of one-dimensional
and two-dimensional electrophoresis. The results showed that trichloroacetic acid/acetone extraction methods with
two different concentrations of 2-mercaptoethanol produced the highest protein concentration and yield for both adult
and larva tissues, respectively. Meanwhile, trichloroacetic acid/acetone with dithiothreitol extraction method gave
better separation of spots and intensity for both larva and adult tissues compared to other methods tested. As such, we
concluded that trichloroacetic acid/acetone with dithiothreitol successfully yielded high total protein concentration and
good separation of two-dimensional electrophoresis gel spots in both adult and larva P. xylostella.

Matched MeSH terms: Electrophoresis, Gel, Two-Dimensional
Pulsed-field gel electrophoresis (PFGE): A review of the "gold standard" for bacteria typing and current alternatives

Neoh HM, Tan XE, Sapri HF, Tan TL

Infect Genet Evol, 2019 10;74:103935.
PMID: 31233781 DOI: 10.1016/j.meegid.2019.103935

Pulsed-field gel electrophoresis (PFGE) is considered the "gold standard" for bacteria typing. The method involves enzyme restriction of bacteria DNA, separation of the restricted DNA bands using a pulsed-field electrophoresis chamber, followed by clonal assignment of bacteria based on PFGE banding patterns. Various PFGE protocols have been developed for typing different bacteria, leading it to be one of the most widely used methods for phylogenetic studies, food safety surveillance, infection control and outbreak investigations. On the other hand, as PFGE is lengthy and labourious, several PCR-based typing methods can be used as alternatives for research purposes. Recently, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) and whole genome sequencing (WGS) have also been proposed for bacteria typing. In fact, as WGS provides more information, such as antimicrobial resistance and virulence of the tested bacteria in comparison to PFGE, more and more laboratories are currently transitioning from PFGE to WGS for bacteria typing. Nevertheless, PFGE will remain an affordable and relevant technique for small laboratories and hospitals in years to come.

Matched MeSH terms: Electrophoresis, Gel, Pulsed-Field
Fulltext Agarose-resolvable InDel markers based on whole genome re-sequencing in cucumber

Adedze YMN, Lu X, Xia Y, Sun Q, Nchongboh CG, Alam MA, et al.

Sci Rep, 2021 02 16;11(1):3872.
PMID: 33594240 DOI: 10.1038/s41598-021-83313-x

Insertion and Deletion (InDel) are common features in genomes and are associated with genetic variation. The whole-genome re-sequencing data from two parents (X1 and X2) of the elite cucumber (Cucumis sativus) hybrid variety Lvmei No.1 was used for genome-wide InDel polymorphisms analysis. Obtained sequence reads were mapped to the genome reference sequence of Chinese fresh market type inbred line '9930' and gaps conforming to InDel were pinpointed. Further, the level of cross-parents polymorphism among five pairs of cucumber breeding parents and their corresponding hybrid varieties were used for evaluating hybrid seeds purity test efficiency of InDel markers. A panel of 48 cucumber breeding lines was utilized for PCR amplification versatility and phylogenetic analysis of these markers. In total, 10,470 candidate InDel markers were identified for X1 and X2. Among these, 385 markers with more than 30 nucleotide difference were arbitrary chosen. These markers were selected for experimental resolvability through electrophoresis on an Agarose gel. Two hundred and eleven (211) accounting for 54.81% of markers could be validated as single and clear polymorphic pattern while 174 (45.19%) showed unclear or monomorphic genetic bands between X1 and X2. Cross-parents polymorphism evaluation recorded 68 (32.23%) of these markers, which were designated as cross-parents transferable (CPT) InDel markers. Interestingly, the marker InDel114 presented experimental transferability between cucumber and melon. A panel of 48 cucumber breeding lines including parents of Lvmei No. 1 subjected to PCR amplification versatility using CPT InDel markers successfully clustered them into fruit and common cucumber varieties based on phylogenetic analysis. It is worth noting that 16 of these markers were predominately associated to enzymatic activities in cucumber. These agarose-based InDel markers could constitute a valuable resource for hybrid seeds purity testing, germplasm classification and marker-assisted breeding in cucumber.

Matched MeSH terms: Electrophoresis, Agar Gel
Fulltext Extraction of gelatin from salmon (Salmo salar) fish skin using trypsin-aided process: optimization by Plackett-Burman and response surface methodological approaches

Fan H, Dumont MJ, Simpson BK

J Food Sci Technol, 2017 Nov;54(12):4000-4008.
PMID: 29085142 DOI: 10.1007/s13197-017-2864-5

Gelatin from salmon (Salmo salar) skin with high molecular weight protein chains (α-chains) was extracted using trypsin-aided process. Response surface methodology was used to optimise the extraction parameters. Yield, hydroxyproline content and protein electrophoretic profile via sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of gelatin were used as responses in the optimization study. The optimum conditions were determined as: trypsin concentration at 1.49 U/g; extraction temperature at 45 °C; and extraction time at 6 h 16 min. This response surface optimized model was significant and produced an experimental value (202.04 ± 8.64%) in good agreement with the predicted value (204.19%). Twofold higher yields of gelatin with high molecular weight protein chains were achieved in the optimized process with trypsin treatment when compared to the process without trypsin.

Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
Fulltext Biochemical properties of red tilapia (Oreochromis niloticus) protein hydrolysates

Shamloo, M., Bakar, J., Mat Hashim, D., Khatib, A.

International Food Research Journal, 2012;19(1):183-188.
MyJurnal

The amino-acid composition, 2, 2-Diphenyl-1-picryhydrazyl (DPPH) radical-scavenging activity, and peptide patterns of tilapia protein hydrolysates produced by the enzymatic hydrolysis of Alcalase (AH), Flavourzyme (FH) and Protamex (PH) for 5h using pH-stat method were studied. The ratio of essential amino acids to non-essential amino acids increased after hydrolysis in all samples; however, no significant differences among them were observed. AH had a highest (P < 0.05) DPPH radical-scavenging activity, but no significant difference in the DPPH between FH and PH was observed. SDS-PAGE patterns for all the hydrolysates showed significant (P < 0.05) reduction in the number and the intensity of the bands with increasing time of hydrolysis. Flavourzyme showed the lowest rate of hydrolytic activity towards the tilapia mince.

Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
Fulltext The spectrum of beta-globin gene mutations in children with beta-thalassaemia major from Kota Kinabalu, Sabah, Malaysia

Thong MK, Soo TL

Singapore Med J, 2005 Jul;46(7):340-3.
PMID: 15968446

Beta-thalassaemia major is one of the commonest genetic disorders in South East Asia. The strategy for the community control of beta-thalassaemia major requires the characterisation of the spectrum of beta-globin gene mutations in any multi-ethnic population. There is only a single report of mutation analyses of the beta-globin gene in an isolated Kadazandusun community in Kota Belud, Sabah, Malaysia, which showed the presence of a common 45 kb deletion.

Matched MeSH terms: Electrophoresis, Gel, Two-Dimensional
Identification of major and cross-reactive allergens of local freshwater snail (Pila polita) and the impact of thermal and non-thermal food processing on allergen stability

Misnan R, Kamarazaman NA, Sockalingam K, Yadzir ZHM, Bakhtiar F, Abdullah N, et al.

J Sci Food Agric, 2023 Sep;103(12):5819-5830.
PMID: 37092326 DOI: 10.1002/jsfa.12659

BACKGROUND: Snail allergy is rare but can be fatal. Pila polita, a freshwater snail, was considered as a popular exotic food, particularly in tropical countries, and consumed in processed forms. Thus, the purpose of this study was to identify the major and cross-reactive allergens of P. polita and to determine the impact of food processing on the allergen stability.
RESULTS: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis fractionated raw snail extract to approximately 24 protein bands, between 9 and 245 kDa. The prominent band at 33 kDa was detected in all raw and processed snail extracts. Immunoblotting tests of the raw extract demonstrated 19 immunoglobulin E (IgE)-binding proteins, and four of them, at 30, 35, 42 and 49 kDa, were revealed as the major IgE-binding proteins of P. polita. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identified the 49 and 42 kDa major allergens as actin, whereas the 30 and 35 kDa major allergens were identified as tropomyosin. Immunoblotting revealed that the raw snail had more allergenic proteins than the processed snail. The degree of allergenicity in decreasing order was raw > brine pickled> boiled > roasted > fried > vinegar pickled. The presence of cross-reactivity between P. polita and the shellfish tested was exhibited with either no, complete, or partial inhibitions.
CONCLUSION: Actin and tropomyosin were identified as the major and cross-reactive allergens of P. polita among local patients with snail allergy. Those major allergens are highly stable to high temperatures, acidic pH, and high salt, which might played a crucial role in snail allergy in Malaysia. © 2023 Society of Chemical Industry.

Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
Fruit-specific expression of papaya subtilase gene

Othman R, Nuraziyan A

J Plant Physiol, 2010 Jan 15;167(2):131-7.
PMID: 19729222 DOI: 10.1016/j.jplph.2009.07.015

Subtilisin-like serine proteases (EC 3.4.21) consist of a widespread family of enzymes that is involved in various processes including in plants. The full-length cDNA (CpSUB1) and the corresponding genomic DNA for papaya subtilase have been obtained using rapid amplification of cDNA ends (RACEs) and PCR primer walking techniques, respectively. The cDNA clone contains an open reading frame of 2316bp encoding 772 amino acids with a calculated molecular mass of 82.6kDa and an isoelectric point (pI) of 8.97. The CpSUB1 gene is composed of nine exons and eight introns. The amino acid sequence encoded by CpSUB1 shared high identity (>60%) with the amino acid sequence of other plant subtilisin-like proteases. Sequence analysis of CpSUB1 revealed the presence of a possible signal peptide (25 amino acid residues) and an NH(2)-terminal prosequence (88 amino acid residues). In addition, papaya subtilase possesses the characteristic subtilisin catalytic triad amino acids namely Asp, His and Ser, together with the substrate-binding site, Asn. DNA hybridization analysis showed that subtilase gene exists as a single copy in the papaya genome. RNA hybridization analyses showed that expression of the subtilase transcripts was only detected in mesocarp but not in non-fruit tissues. Gene expression in fruit tissues reached the highest level during the ripening stage at which the fruits undergo dramatic softening process. Subsequently, pro-subtilase ( approximately 80kDa) was expressed as recombinant pro-enzyme ( approximately 97kDa), which was used to generate antiserum against papaya subtilase, anti-sub. Protein gel blot analysis using anti-sub towards total protein extracted from all ripening stages revealed that a protein with a molecular mass of approximately 70kDa reacted with the antiserum. Hence both RNA hybridization and protein gel blot analyses confirmed the presence of subtilase during papaya fruit ripening, pointing to its possible involvement in this important process.

Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
Population genetics of allozyme variation in Neurospora intermedia

Spieth PT

Genetics, 1975 Aug;80(4):785-805.
PMID: 1193373

Electrophoretically detectable variation in the fungus Neurospora intermedia has been surveyed among isolates from natural populations in Malaya, Papua, Australia and Florida. The principal result is a pattern of genetic variation within and between populations that is qualitatively no different than the well documented patterns for Drosophila and humans. In particular, there is a high level of genetic variation, the majority of which occurs at the level of local populations. Evidence is presented which argues that N. intermedia has a population structure analogous to that of an annual vascular plant with a high level of vegetative reproduction. Sexual reproduction appears to be a regular feature in the biology of the species. Substantial heterokaryon function seems unlikely in natural populations of N. intermedia. Theoretical considerations concerning the mechanisms underlying the observed pattern of variation most likely should be consistent with haploid selection theory. The implications of this constraint upon the theory are discussed in detail, leading to the presentation of a model based upon the concept of environmental heterogenicity. The essence of the model, which is equally applicable to haploid and diploid situations, is a shifting distribution of multiple adaptive niches among local populations such that a given population has a small net selective pressure in favor of one allele or another, depending upon its particular distribution of niches. Gene flow among neighboring populations with differing net selective pressures is postulated as the principal factor underlying intrapopulational allozyme variation.

Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
Fulltext Enzyme Hydrolysates from Stichopus horrens as a New Source for Angiotensin-Converting Enzyme Inhibitory Peptides

Forghani B, Ebrahimpour A, Bakar J, Abdul Hamid A, Hassan Z, Saari N

Evid Based Complement Alternat Med, 2012;2012:236384.
PMID: 22927875 DOI: 10.1155/2012/236384

Stichopus horrens flesh was explored as a potential source for generating peptides with angiotensin-converting enzyme (ACE) inhibitory capacity using 6 proteases, namely alcalase, flavourzyme, trypsin, papain, bromelain, and protamex. Degree of hydrolysis (DH) and peptide profiling (SDS-PAGE) of Stichopus horrens hydrolysates (SHHs) was also assessed. Alcalase hydrolysate showed the highest DH value (39.8%) followed by flavourzyme hydrolysate (32.7%). Overall, alcalase hydrolysate exhibited the highest ACE inhibitory activity (IC(50) value of 0.41 mg/mL) followed by flavourzyme hydrolysate (IC(50) value of 2.24 mg/mL), trypsin hydrolysate (IC(50) value of 2.28 mg/mL), papain hydrolysate (IC(50) value of 2.48 mg/mL), bromelain hydrolysate (IC(50) value of 4.21 mg/mL), and protamex hydrolysate (IC(50) value of 6.38 mg/mL). The SDS-PAGE results showed that alcalase hydrolysate represented a unique pattern compared to others, which yielded potent ACE inhibitory peptides with molecular weight distribution lower than 20 kDa. The evaluation of the relationship between DH and IC(50) values of alcalase and flavourzyme hydrolysates revealed that the trend between those parameters was related to the type of the protease used. We concluded that the tested SHHs would be used as a potential source of functional ACE inhibitory peptides for physiological benefits.

Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
Genetic dynamics of Salmonella typhi--diversity in clonality

Pang T

Trends Microbiol, 1998 Sep;6(9):339-42.
PMID: 9778724
Matched MeSH terms: Electrophoresis, Gel, Pulsed-Field
Fulltext High performance liquid chromatography (HPLC) as a screening tool for classical Beta-thalassaemia trait in malaysia

George E, Jamal AR, Khalid F, Osman KA

Malays J Med Sci, 2001 Jul;8(2):40-6.
PMID: 22893759 MyJurnal

Beta-thalassaemia is characterized by a decrease (β(+)) or absence (β(0)) in the synthesis of β-globin chains of human haemoglobin. The heterozygous state for β(+) or β(0) result in β-thalassaemia trait in which the hallmark is the presence of an elevated level of Haemoglobin (Hb) A(2) (α(2)δ(2)). In the past, the traditional methods such as cellulose acetate electrophoresis with elution and microcolumn chromatrography have been the techniques used by the majority of the laboratories in Malaysia for the estimation of (Hb) A(2) levels. The recommended method currently is high performance liquid chromatography which has only been introduced in a few laboratories in the country.

Matched MeSH terms: Electrophoresis, Cellulose Acetate
Fulltext Thalassemia among blood donors at the Hospital Universiti Sains Malaysia

Rosline H, Ahmed SA, Al-Joudi FS, Rapiaah M, Naing NN, Adam NA

Southeast Asian J Trop Med Public Health, 2006 May;37(3):549-52.
PMID: 17120978

The aim of this study was to screen and identify the types of thalassemia among blood donors at the Hospital Universiti Sains Malaysia (HUSM). Thalassemia screening was performed by hemoglobin electrophoresis. A total number of 80 blood samples were obtained from donors at the Transfusion Medicine Unit, HUSM. The ethnic origins of the donors were Malays (n=73, 91.3%) and non-Malays (n=7, 8.75%). Males comprised 88.1% of the donors. Thalassemia was detected in 16.25% (n=13) of the blood donors. Of those with thalassemia, 46.2% (6/13) were anemic. Microcytosis and hypochromia were detected in 84.6% (n=l1) and 84.6% (n=l1) of these donors, respectively. The types of thalassemias detected were Hb E, 11.25% (n=9/80) and beta thalassemia trait, 5% (n=4/80). Among the thalassemias detected, the Hb E hemoglobinopathy was comprised of Hb E/ alpha-thalassemia (38.5%: n=5), Hb E /beta-thalassemia (23.1%: n=3), Hb E trait (7.6%: n=1) and beta-thalassemia (30.8%: n=4). In conclusion, screening for thalassemia trait should be included as part of a standard blood testing before blood donation. Further studies are required to look at the effects of donated thalassemic blood.

Matched MeSH terms: Electrophoresis
Genetic microdifferentiation in the Semai Senoi of Malaysia

Fix AG, Lie-injo LE

Am J Phys Anthropol, 1975 Jul;43(1):47-55.
PMID: 1155591

Blood samples, demographic and cultural data were collected from seven settlements of Semai Senoi, a swidden farming ethnic group of Malaysia. Three genetic loci (ABO blood group, hereditary ovalcytosis, and hemoglobin) were analyzed in a total sample of 546 individuals. These data indicate a considerable degree of genetic microdifferentiation in this area of the Semai distribution. Parent-offspring birthplace data (analyzed by means of a migration matrix) and settlement histories show that settlements are not strongly isolated. Genetic differences in the study area demonstrate a reasonable correspondence with migration and the history of the settlements. Genetic convergence also occurs through the addition of migrant groups to established populations leading to new patterns of marriage between donor and recipient groups. The genetic structure of the total Semai population through time thus comprises a mosaic of shifiting allele frequencies in a series of semi-isolated local populations.

Matched MeSH terms: Electrophoresis, Starch Gel
Multiple fetal and adult hemoglobins in Malaysian Macaca nemestrina: consequences of a-chain variation

Nute PE, Pataryas HA

Am J Phys Anthropol, 1974 Jan;40(1):17-25.
PMID: 4206325
Matched MeSH terms: Electrophoresis, Starch Gel
Gelsolin and ceruloplasmin as potential predictive biomarkers for cervical cancer by 2D-DIGE proteomics analysis

Lokamani I, Looi ML, Md Ali SA, Mohd Dali AZ, Ahmad Annuar MA, Jamal R

Pathol Oncol Res, 2014 Jan;20(1):119-29.
PMID: 23925487 DOI: 10.1007/s12253-013-9670-9

This study aimed to identify candidate proteins which may serve as potential biological markers for cervical cancer using 2D-DIGE. Serum samples of controls, patients with cervical intraepithelial neoplasia grade 3 (CIN 3), squamous cell carcinoma of early (SCC I and II) and late (SCC III and IV) stage were subjected to 2D-DIGE. Differentially expressed spots were identified by tandem mass spectrometry. Validation of candidate proteins in serum and tissue samples were then performed by ELISA and immunohistochemistry (IHC) analysis respectively. A total of 20 differentially expressed proteins were identified. These proteins were found to play key roles in the apoptosis pathway, complement system, various types of transportation such as hormones, fatty acids, lipid, vitamin E and drug transportation, coagulation cascade, regulation of iron and immunologic response. Based on their functional relevancy to the progression of various cancers, 4 proteins namely the complement factor H, CD5-like antigen, gelsolin and ceruloplasmin were chosen for further validation using ELISA. Biological network analysis showed that ceruloplasmin and gelsolin are closely interacted with the oncogene NF-κb. These two proteins were further validated using the IHC. Gelsolin and ceruloplasmin may serve as potential predictive biomarkers for the progression of high grade lesions.

Matched MeSH terms: Two-Dimensional Difference Gel Electrophoresis/methods
Fulltext High genetic diversity of Enterococcus faecium and Enterococcus faecalis clinical isolates by pulsed-field gel electrophoresis and multilocus sequence typing from a hospital in Malaysia

Weng PL, Ramli R, Shamsudin MN, Cheah YK, Hamat RA

Biomed Res Int, 2013;2013:938937.
PMID: 23819125 DOI: 10.1155/2013/938937

Little is known on the genetic relatedness and potential dissemination of particular enterococcal clones in Malaysia. We studied the antibiotic susceptibility profiles of Enterococcus faecium and Enterococcus faecalis and subjected them to pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). E. faecium and E. faecalis displayed 27 and 30 pulsotypes, respectively, and 10 representative E. faecium and E. faecalis isolates (five each) yielded few different sequence types (STs): ST17 (2 isolates), ST78, ST203, and ST601 for E. faecium, and ST6, ST16, ST28, ST179, and ST399 for E. faecalis. Resistance to tazobactam-piperacillin and ampicillin amongst E. faecium isolates was highly observed as compared to E. faecalis isolates. All of the isolates were sensitive to vancomycin and teicoplanin. The presence of epidemic and nosocomial strains of selected E. faecium STs: 17, 78, and 203 and E. faecalis ST6 as well as high rates of resistance to multiple antibiotics amongst E. faecium isolates is of a particular concern.

Matched MeSH terms: Electrophoresis, Gel, Pulsed-Field/methods*
Capillary electrophoresis with capacitively coupled contactless conductivity detection for the determination of cis/trans isomers of octadec-9-enoic acid and other long chain fatty acids

Wong YF, Saad B, Makahleh A

J Chromatogr A, 2013 May 17;1290:82-90.
PMID: 23578483 DOI: 10.1016/j.chroma.2013.03.014

A capillary electrophoresis (CE)-capacitively coupled contactless conductivity detection (C(4)D) method for the simultaneous separation of eleven underivatized fatty acids (FAs), namely, lauric, myristic, tridecanoic (internal standard), pentadecanoic, palmitic, stearic, oleic, elaidic, linoleic, linolenic and arachidic acids is described. The separation was carried out in normal polarity mode at 20 °C, 30 kV and using hydrodynamic injection (50 mbar for 1 s). The separation was achieved in a bare fused-silica capillary (70 cm × 75 μm i.d.) using a background electrolyte of methyl-β-cyclodextrin (~6 mM) and heptakis-(2,3,6-tri-O-methyl)-β-cyclodextrin (~8 mM) dissolved in a mixture of Na2HPO4/KH2PO4 (5 mM, pH 7.4):ACN:MeOH:n-octanol (3:4:2.5:0.5, v/v/v/v). C(4)D parameters were set at fixed amplitude of 100 V and frequency of 1000 kHz. The developed method was validated. Calibration curves of the ten FAs were well correlated (r(2)>0.99) within the range of 5-250 μg mL(-1) for lauric acid, and 3-250 μg mL(-1) for the other FAs. The method was simple and sensitive with detection limits (S/N=3) of 0.9-1.9 μg mL(-1) and good relative standard deviations of intra- and inter-day for migration times and peak areas (≤9.7%) were achieved. The method was applied to the determination of FAs in margarine samples. The proposed method offers distinct advantages over the GC and HPLC methods, especially in terms of simplicity (without derivatization) and sensitivity.

Matched MeSH terms: Electrophoresis, Capillary/methods*
Fulltext Identification of actinomycete communities in Antarctic soil from Barrientos Island using PCR-denaturing gradient gel electrophoresis

Learn-Han L, Yoke-Kqueen C, Shiran MS, Vui-Ling CM, Nurul-Syakima AM, Son R, et al.

Genet. Mol. Res., 2012;11(1):277-91.
PMID: 22370930 DOI: 10.4238/2012.February.8.3

The diversity of specific bacteria taxa, such as the actinomycetes, has not been reported from the Antarctic island of Barrientos. The diversity of actinomycetes was estimated with two different strategies that use PCR-denaturing gradient gel electrophoresis. First, a PCR was applied, using a group-specific primer that allows selective amplification of actinomycete sequences. Second, a nested-PCR approach was used that allows the estimation of the relative abundance of actinomycetes within the bacterial community. Molecular identification, which was based on 16S rDNA sequence analysis, revealed eight genera of actinomycetes, Actinobacterium, Actinomyces, an uncultured Actinomycete, Streptomyces, Leifsonia, Frankineae, Rhodococcus, and Mycobacterium. The uncultured Actinomyces sp and Rhodococcus sp appear to be the prominent genera of actinomycetes in Barrientos Island soil. PCR-denaturing gradient gel electrophoresis patterns were used to look for correlations between actinomycete abundance and environmental characteristics, such as type of rookery and vegetation. There was a significant positive correlation between type of rookery and abundance of actinomycetes; soil samples collected from active chinstrap penguin rookeries had the highest actinomycete abundance. Vegetation type, such as moss, which could provide a microhabitat for bacteria, did not correlate significantly with actinomycete abundance.

Matched MeSH terms: Denaturing Gradient Gel Electrophoresis/methods

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