Displaying publications 1 - 20 of 294 in total

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  1. Long-Range Polymerase Chain Reaction
    Kee PS, Karunanathie H, Maggo SDS, Kennedy MA, Chua EW
    Methods Mol Biol, 2023;2967:181-192.
    PMID: 37608112 DOI: 10.1007/978-1-0716-3358-8_15
    Polymerase chain reaction (PCR) is a laboratory technique used to amplify a targeted region of DNA, demarcated by a set of oligonucleotide primers. Long-range PCR is a form of PCR optimized to facilitate the amplification of large fragments. Using the adapted long-range PCR protocol described in this chapter, we were able to generate PCR products of 6.6, 7.2, 13, and 20 kb from human genomic DNA samples. For some of the long PCRs, successful amplification was not possible without the use of PCR enhancers. Thus, we also evaluated the impact of some enhancers on long-range PCR and included the findings as part of this updated chapter.
    Matched MeSH terms: DNA Primers/genetics
  2. Fulltext Pengimejan resonans magnet kefungsian: pemerolehan, analisis dan pentafsiran data
    Ahmad Nazlim Yusoff, Mohd Harith Hashim, Mohd Mahadir Ayob, Iskandar Kassim
    Jurnal Sains Kesihatan Malaysia, 2005;3(2):19-37.
    MyJurnal
    Kajian garis pangkal pengimejan resonans magnet kefungsian (fMRI) telah dijalankan di Jabatan Radiologi, Hospital Universiti Kebangsaan Malaysia ke atas seorang subjek lelaki sihat berumur 25 tahun menggunakan sistem pengimejan resonans magnet (MRI) 1.5 T. Kajian ini menggunakan gerakan jari tangan kanan dan kiri untuk merangsang aktiviti neuron di dalam korteks serebrum. Subjek diarahkan supaya menekan jari-jari pada ibu jari secara bergilir-gilir semasa imbasan kefungsian dilakukan. Paradigma 5 kitar aktifrehat digunakan dengan setiap kitar masing-masing mengandungi 20 siri pengukuran. Keputusan menunjukkan bahawa rantau otak yang aktif akibat gerakan jari adalah girus presentral merangkumi kawasan motor primer. Pengaktifan otak adalah secara kontralateral terhadap gerakan jari tangan kanan dan kiri. Keamatan isyarat keadaan aktif didapati lebih tinggi daripada keamatan isyarat keadaan rehat. Analisis yang dilakukan ke atas beberapa rantau pengaktifan yang diminati (ROI) pada beberapa hirisan menunjukkan perbezaan yang bererti (p < 0.05) antara keamatan keadaan aktif dan rehat untuk nilai ambang statistik (Z) = 1.0 dan 1.5. Perbezaan purata antara kedua-dua purata keamatan isyarat keadaan aktif dan rehat pada manamana hirisan untuk kedua-dua nilai Z menunjukkan magnitud pengaktifan yang lebih tinggi pada hemisfera kanan otak iaitu apabila subjek menggerakkan tangan kirinya. Bilangan voksel yang aktif juga didapati lebih tinggi pada hemisfera kanan berbanding pada hemisfera kiri otak. Keputusan ini menyokong fakta bahawa bagi subjek yang tidak kidal, kawasan pengaktifan motor pada hemisfera kanan otak semasa gerakan jari tangan kiri mengalami rangsangan hemodinamik yang lebih tinggi berbanding dengan hemisfera kiri otak semasa gerakan jari tangan kanan. Fenomena rangsangan hemodinamik yang diperhatikan dalam kajian ini dibincangkan berdasarkan kepada kebergantungan kontras isyarat kepada aras oksigen darah (BOLD).
    Matched MeSH terms: DNA Primers
  3. Fulltext Wolbachia infection in field-collected Aedes aegypti in Yunnan Province, southwestern China
    Zhang H, Gao J, Ma Z, Liu Y, Wang G, Liu Q, et al.
    Front Cell Infect Microbiol, 2022;12:1082809.
    PMID: 36530420 DOI: 10.3389/fcimb.2022.1082809
    BACKGROUND: Wolbachia is gram-negative and common intracellular bacteria, which is maternally inherited endosymbionts and could expand their propagation in host populations by means of various manipulations. Recent reports reveal the natural infection of Wolbachia in Aedes Aegypti in Malaysia, India, Philippines, Thailand and the United States. At present, none of Wolbachia natural infection in Ae. aegypti has been reported in China.

    METHODS: A total of 480 Ae. aegypti adult mosquitoes were collected from October and November 2018 based on the results of previous investigations and the distribution of Ae. aegypti in Yunnan. Each individual sample was processed and screened for the presence of Wolbachia by PCR with wsp primers. Phylogenetic trees for the wsp gene was constructed using the neighbour-joining method with 1,000 bootstrap replicates, and the p-distance distribution model of molecular evolution was applied.

    RESULTS: 24 individual adult mosquito samples and 10 sample sites were positive for Wolbachia infection. The Wolbachia infection rate (IR) of each population ranged from 0 - 41.7%. The infection rate of group A alone was 0%-10%, the infection rate of group B alone was 0%-7.7%, and the infection rate of co-infection with A and B was 0-33.3%.

    CONCLUSIONS: Wolbachia infection in wild Ae. aegypti in China is the first report based on PCR amplification of the Wolbachia wsp gene. The Wolbachia infection is 5%, and the wAlbA and wAlbB strains were found to be prevalent in the natural population of Ae. aegypti in Yunnan Province.

    Matched MeSH terms: DNA Primers
  4. A transposon-based genetic marker for conspecific identity within the Bactrocera dorsalis species complex
    Zimowska GJ, Xavier N, Qadri M, Handler AM
    Sci Rep, 2024 Jan 22;14(1):1924.
    PMID: 38253542 DOI: 10.1038/s41598-023-51068-2
    Here we describe a molecular approach to assess conspecific identity that relies on the comparison of an evolved mutated transposable element sequence and its genomic insertion site in individuals from closely related species. This was explored with the IFP2 piggyBac transposon, originally discovered in Trichoplusia ni as a 2472 bp functional element, that was subsequently found as mutated elements in seven species within the Bactrocera dorsalis species complex. In a B. dorsalis [Hendel] strain collected in Kahuku, Hawaii, a degenerate 2420 bp piggyBac sequence (pBacBd-Kah) having ~ 94.5% sequence identity to IFP2 was isolated, and it was reasoned that common species, or strains within species, should share the same evolved element and its precise genomic insertion site. To test this assumption, PCR using primers to pBacBd-Kah and adjacent genomic sequences was used to isolate and compare homologous sequences in strains of four sibling species within the complex. Three of these taxa, B. papayae, B. philippinensis, and B. invadens, were previously synonymized with B. dorsalis, and found to share nearly identical pBacBd-Kah homologous elements (> 99% nucleotide identity) within the identical insertion site consistent with conspecific species. The fourth species tested, B. carambolae, considered to be a closely related yet independent species sympatric with B. dorsalis, also shared the pBacBd-Kah sequence and insertion site in one strain from Suriname, while another divergent pBacBd-Kah derivative, closer in identity to IFP2, was found in individuals from French Guiana, Bangladesh and Malaysia. This data, along with the absence of pBacBd-Kah in distantly related Bactrocera, indicates that mutated descendants of piggyBac, as well as other invasive mobile elements, could be reliable genomic markers for common species identity.
    Matched MeSH terms: DNA Primers
  5. Fulltext Isolation and characterization of novel microsatellite markers in commercial selected golden Malaysian arowana fish, Scleropages formosus (Osteoglossidae)
    Manoharan B, Sulaimen Z, Omar F, Othman RY, Mohamed SZ, Bhassu S
    Genet. Mol. Res., 2011;10(2):712-6.
    PMID: 21523650 DOI: 10.4238/vol10-2gmr944
    Malaysian arowana (dragonfish; Scleropages formosus) is an ancient osteoglossid fish from southeast Asia. Due to the high demand of the ornamental fish trade and because of habitat loss, the species is close to extinction. We isolated and characterized 10 polymorphic microsatellites of this species, using 5'-anchored PCR. The number of alleles at the 10 microsatellite loci ranged from 2 to 28, with a mean of 7.8/locus. The observed heterozygosity ranged from 0.03 to 0.93 (mean: 0.39), whereas the expected heterozygosity ranged from 0.03 to 0.94 (mean: 0.46). Seven microsatellites deviated from Hardy-Weinberg equilibrium, and three conformed to Hardy-Weinberg equilibrium and were in linkage equilibrium. These 10 novel microsatellites should facilitate studies of genetic diversity and population structure of arowana to help plan actions for the conservation of the indigenous Malaysian arowana.
    Matched MeSH terms: DNA Primers/genetics*
  6. Isolation by the 5' anchored PCR technique and characterization of eighteen microsatellite loci in horseshoe crab (Tachypleus gigas)
    Ling LP, Adibah AB, Tan SG, Christianus A, Faridah QZ
    J Genet, 2011 Dec;90(3):e101-4.
    PMID: 22232191
    Matched MeSH terms: DNA Primers/chemistry
  7. Fulltext A duplex endpoint PCR assay for rapid detection and differentiation of Leptospira strains
    Benacer D, Zain SNM, Lewis JW, Khalid MKNM, Thong KL
    Rev Soc Bras Med Trop, 2017 Mar-Apr;50(2):239-242.
    PMID: 28562762 DOI: 10.1590/0037-8682-0364-2016
    INTRODUCTION:: This study aimed to develop a duplex endpoint PCR assay for rapid detection and differentiation of Leptospira strains.

    METHODS:: Primers were designed to target the rrs (LG1/LG2) and ligB (LP1/LP2) genes to confirm the presence of the Leptospira genus and the pathogenic species, respectively.

    RESULTS:: The assay showed 100% specificity against 17 Leptospira strains with a limit of detection of 23.1pg/µl of leptospiral DNA and sensitivity of 103 leptospires/ml in both spiked urine and water.

    CONCLUSIONS:: Our duplex endpoint PCR assay is suitable for rapid early detection of Leptospira with high sensitivity and specificity.
    Matched MeSH terms: DNA Primers*
  8. Fulltext Rapid Detection of Porcine DNA in Meatball Using Recombinase Polymerase Amplification Couple with Lateral Flow Immunoassay for Halal Authentication
    Yusop MHM, Bakar MFA, Kamarudin KR, Mokhtar NFK, Hossain MAM, Johan MR, et al.
    Molecules, 2022 Nov 22;27(23).
    PMID: 36500215 DOI: 10.3390/molecules27238122
    Point-of-care diagnostic methods for animal species determination are critical for rapid, simple, and accurate enforcement of food labelling. PCR is the most common method for species identification. However, the requirement of using a thermal cycler created drawbacks for the PCR application, particularly in low-resource settings. Hence, in this study, a method for porcine DNA detection using recombinase polymerase amplification (RPA), coupled with nucleic acid lateral flow immunoassay (NALFIA), was developed. Porcine-specific primers targeting pig (Sus scrofa) cytochrome b gene fragments specifically amplify a 197 bp fragment of the mitochondrial gene as being visualized by 2% agarose gel and PCRD NALFIA. The reaction temperature and time were 39 °C and 20 min, respectively. Herein, the specificity of the primers to porcine was confirmed after being assayed against six animal species, namely cow, goat, chicken, duck, dog, and rabbit. The porcine-specific RPA assay shows a high limit of detection of 0.01 ng/µL pork DNA. Based on the preliminary performance data obtained from this study, the potential of this method as a rapid and sensitive tool for porcine DNA detection in meat-based products is foreseen.
    Matched MeSH terms: DNA Primers/genetics
  9. The utility of RAPD markers for the determination of genetic variation in oil palm (Elaeis guineensis)
    Shah FH, Rashid O, Simons AJ, Dunsdon A
    Theor Appl Genet, 1994 Nov;89(6):713-8.
    PMID: 24178016 DOI: 10.1007/BF00223710
    The genetic variation among different accessions of oil-palm germplasm collected from Africa was estimated using random primers and the polymerase chain reaction. The present study revealed high levels of genetic variation in these accessions. Electrophoresis of the amplification products indicated that nine out of 20 primers were able to generate polymorphic products ranging in length from 0.2 kb to 2.3 kb. No individual palm or population-specific products were observed. Greatest diversity was seen in Zaire population 5 and the least in Zaire population 2.
    Matched MeSH terms: DNA Primers
  10. Fulltext Pengaktifan otak akibat gerakan jari bagi subjek dominan tangan kanan dan kiri
    Ahmad Nazlim Yusoff, Mohd Harith Hashim, Mohd Mahadir Ayob, Iskandar Kassim
    Jurnal Sains Kesihatan Malaysia, 2006;4(2):63-83.
    MyJurnal
    Kajian garis pangkal pengimejan resonans magnet kefungsian (fMRI) telah dijalankan ke atas 2 orang subjek lelaki sihat dominan tangan kanan dan kiri. Kajian ini menggunakan gerakan jari tangan kanan dan kiri untuk merangsang aktiviti neuron di dalam korteks serebrum. Subjek diarahkan supaya menekan jari-jari pada ibu jari secara bergilir-gilir semasa imbasan fMRI dilakukan. Paradigma 5 kitar aktif-rehat digunakan dengan setiap kitar mengandungi satu blok aktif dan satu blok rehat dengan 10 siri pengukuran untuk setiap blok. Seratus isipadu imej fMRI bagi setiap subjek dianalisis menggunakan pekej perisian MatLab dan SPM2. Model linear am (GLM) digunakan untuk menganggar secara statistik parameter yang mencirikan model rangsangan hemodinamik bagi gerakan jari. Kesimpulan mengenai pengaktifan otak yang diperhatikan dijana secara statistik berasaskan teori medan rawak (RFT) Gaussian. Keputusan menunjukkan bahawa rantau otak yang aktif akibat gerakan jari adalah pada girus presentral merangkumi kawasan motor primer. Pengaktifan otak adalah secara kontralateral terhadap gerakan jari tangan kanan dan kiri. Keamatan isyarat keadaan aktif didapati lebih tinggi secara bererti (p < 0.001) daripada keamatan isyarat keadaan rehat. Bilangan voksel yang aktif didapati lebih tinggi pada hemisfera otak yang mengawal gerakan jari bagi tangan yang tidak dominan untuk kedua-dua subjek. Keputusan ini menyokong fakta bahawa kawasan pengaktifan motor pada hemisfera otak semasa gerakan jari tangan yang tidak dominan mengalami rangsangan hemodinamik yang lebih tinggi dan kawasan pengaktifan yang lebih luas berbanding dengan kawasan pengaktifan pada hemisfera otak yang mengawal gerakan jari bagi tangan yang dominan.
    Matched MeSH terms: DNA Primers
  11. Genetic characterization of two mahseer species (tor douronensis and tor tambroides) using microsatellite markers from other cyprinids
    Yuzine Esa, Khairul Adha A. Rahim, Siti Shapor Siraj, Muhammad Fadhil Syukri, Siti Khalijah Daud, Ho GC, et al.
    Sains Malaysiana, 2011;40:1087-1095.
    This study examined the genetic characteristics of twenty-six microsatellite primers developed from three cyprinid fishes (Cyprinus carpio Linnaeus, Barbus barbus Linnaeus and Barbonymus gonionotus Bleeker) in two indigenous mahseer. The Tor douronensis Valenciennes were randomly collected from two locations in Sarawak (N=52), while Tor tambroides Bleeker were obtained from Peninsular Malaysia (N=56). A total of ten and twelve primers were successfully amplified producing four and five polymorphic loci in T. douronensis and T. tambroides, respectively. The number of alleles per locus ranging from 2 to 5 in T. douronensis and 2 to 7 in T. tambroides. A significant deviation from Hardy-Weinberg equilibrium (HWE) was observed at three loci (Barb37, Barb59 and Barb62) in one or more populations in T. tambroides while two loci (Barb37 and Barb62) were deviated in T. douronensis population of Batang Ai. Population structure analysis showed low level of inter-population genetic differentiation in both mahseer. Overall, the identified microsatellite loci should be useful in analysing T. douronensis and T. tambroides natural populations.
    Matched MeSH terms: DNA Primers
  12. Elimination of contamination in loop-mediated isothermal amplification assay for detection of human malaria
    Zen LPY, Lai MY, Lau YL
    Trop Biomed, 2020 Dec 01;37(4):1124-1128.
    PMID: 33612764 DOI: 10.47665/tb.37.4.1124
    The LAMP assay, amplifies the target DNA rapidly, with 10-fold greater sensitivity than conventional PCR. The greater sensitivity also comes with greater risks of contamination. To overcome this issue, the current project includes either uracil DNA glycosylase (UDG) or a mineral oil overlay in the LAMP assay. Our results indicated that UDG or a mineral oil overlay can effectively prevent carryover contamination in the LAMP assay for the detection of human malaria. By incorporating these preventative methods, contamination can be eliminated and LAMP can potentially be used in the field; and point of care diagnosis for human malaria.
    Matched MeSH terms: DNA Primers
  13. Development and validation of pan-Leptospira Taqman qPCR for the detection of Leptospira spp. in clinical specimens
    Mohd Ali MR, Mohd Safee AW, Ismail NH, Abu Sapian R, Mat Hussin H, Ismail N, et al.
    Mol Cell Probes, 2018 04;38:1-6.
    PMID: 29524642 DOI: 10.1016/j.mcp.2018.03.001
    BACKGROUND: Early diagnosis of leptospirosis is important for ensuring better clinical management and achieving better outcomes. Currently, serological assays suffer from inconsistent performance and are less useful for early diagnosis of leptospirosis. As an alternative, qPCR is more sensitive, specific and able to detect the presence of leptospiral DNA during the acute phase of the infection. Meanwhile, most molecular assays do not detect the non-pathogenic group of Leptospira, even though these groups may also infect humans, although less frequently and less severely.

    METHODS: A set of primers and probe targeting rrs genes of 22 Leptospira spp. were designed and evaluated on 31 Leptospira isolates, 41 other organisms and 65 clinical samples from suspected patients.

    RESULTS: The developed assay was able to detect as low as 20 fg Leptospira DNA per reaction (equivalent to approximately 4 copies) and showed high specificity against the tested leptospiral strains. No cross amplification was observed with the other organisms. During the evaluation of the confirmed clinical specimens, the developed assay was able to correctly identify all positive samples (n = 10/10). One amplification was observed in a negative sample (n = 1/55). The sequencing of the PCR product of the discordant sample revealed that the sequences were similar to those of L. interrogans and L. kirschneri.

    CONCLUSION: The findings suggest that the developed Taqman qPCR assay is sensitive, specific and has potential to be applied in a larger subsequent study.

    Matched MeSH terms: DNA Primers
  14. Universal mini COI barcode for the identification of fish species in processed products
    Sultana S, Ali ME, Hossain MAM, Asing, Naquiah N, Zaidul ISM
    Food Res Int, 2018 03;105:19-28.
    PMID: 29433207 DOI: 10.1016/j.foodres.2017.10.065
    Species substitution, the use of a low value fish in place of a high value fish, is the biggest problem in international trade and the leading cause of fraud in the fisheries arena sector. Current DNA barcoding systems have partly solved this problem but also failed in many instances to amplify PCR targets from highly processed products because of the degradation of a longer barcode marker (~650bp). In the present study, a novel mini barcode marker (295bp) was developed to discriminate fish species in raw and processed states forms. The barcode primers were cross-tested against 33 fish species and 15 other animal species and found to be universal for all the tested fish varieties. When 20 commercial fish products of five different categories were screened, all commercial fish sample yielded positive bands for the novel fish barcode. PCR product was sequenced to retrieve the species IDs that reflected 55% (11/20) of Malaysian fish products were mislabeled.
    Matched MeSH terms: DNA Primers/genetics; DNA Primers/isolation & purification
  15. Fulltext Characterization of 10 novel microsatellite loci for the brown marbled grouper, Epinephelus fuscoguttatus (Serranidae)
    Mokhtar MA, Normah MN, Kumar SV, Baharum SN
    Genet. Mol. Res., 2011;10(2):885-8.
    PMID: 21644205 DOI: 10.4238/vol10-2gmr1117
    Epinephelus fuscoguttatus is a commercially important marine fish species in southeast Asia. Due to overfishing and water pollution, this species has been declared as near-threatened. Thus, to provide information to help maintain and preserve the species, microsatellites were developed, using an enriched genomic library method. Thirty individuals were collected from the hatchery of the Fishery Research Institute, Terengganu, Malaysia. These individuals, from four to six years old, originated from Sabah and are maintained in captive culture as broodstock. Genomic DNA was extracted from the fins of selected individuals that weighed 3-8 kg. Ten microsatellite loci were found to be polymorphic in this population, with 5 to 21 alleles per locus. Observed and expected heterozygosities ranged from 0.53 to 0.97 and 0.59 to 0.95, respectively. Only one locus deviated significantly from Hardy-Weinberg equilibrium and no significant linkage disequilibrium was found among the pairs of loci. These polymorphic microsatellite loci will be used by the Malaysian Fishery Research Institute for investigating genetic diversity and for developing breeding strategies.
    Matched MeSH terms: DNA Primers
  16. Fulltext Host specialization and phylogenetic diversity of Corynespora cassiicola
    Dixon LJ, Schlub RL, Pernezny K, Datnoff LE
    Phytopathology, 2009 Sep;99(9):1015-27.
    PMID: 19671003 DOI: 10.1094/PHYTO-99-9-1015
    The fungus Corynespora cassiicola is primarily found in the tropics and subtropics, and is widely diverse in substrate utilization and host association. Isolate characterization within C. cassiicola was undertaken to investigate how genetic diversity correlates with host specificity, growth rate, and geographic distribution. C. cassiicola isolates were collected from 68 different plant species in American Samoa, Brazil, Malaysia, and Micronesia, and Florida, Mississippi, and Tennessee within the United States. Phylogenetic analyses using four loci were performed with 143 Corynespora spp. isolates, including outgroup taxa obtained from culture collections: C. citricola, C. melongenae, C. olivacea, C. proliferata, C. sesamum, and C. smithii. Phylogenetic trees were congruent from the ribosomal DNA internal transcribed spacer region, two random hypervariable loci (caa5 and ga4), and the actin-encoding locus act1, indicating a lack of recombination within the species and asexual propagation. Fifty isolates were tested for pathogenicity on eight known C. cassiicola crop hosts: basil, bean, cowpea, cucumber, papaya, soybean, sweet potato, and tomato. Pathogenicity profiles ranged from one to four hosts, with cucumber appearing in 14 of the 16 profiles. Bootstrap analyses and Bayesian posterior probability values identified six statistically significant phylogenetic lineages. The six phylogenetic lineages correlated with host of origin, pathogenicity, and growth rate but not with geographic location. Common fungal genotypes were widely distributed geographically, indicating long-distance and global dispersal of clonal lineages. This research reveals an abundance of previously unrecognized genetic diversity within the species and provides evidence for host specialization on papaya.
    Matched MeSH terms: DNA Primers
  17. Isolation and characterization of microsatellite loci in the Malaysian giant freshwater prawn, Macrobrachium rosenbergii
    Bhassu S, See LM, Hassan R, Siraj SS, Tan SG
    Mol Ecol Resour, 2008 Sep;8(5):983-5.
    PMID: 21585948 DOI: 10.1111/j.1755-0998.2008.02127.x
    Eight single locus microsatellite markers were developed to characterize the Malaysian giant freshwater prawn, Macrobrachium rosenbergii. These microsatellites were isolated from an enriched genomic library contained by using a 5'-anchored polymerase chain reaction technique. Primers were designed to flank the repeat sequences and subsequently used to characterize 30 unrelated individuals of the giant freshwater prawn. The polymerase chain reaction amplification products of these eight microsatellite loci were polymorphic with the number of alleles ranging from two to 10 alleles per locus while the levels of heterozygosity ranged from 0.6333 to 0.8667.
    Matched MeSH terms: DNA Primers
  18. Discovery of a new polyhydroxyalkanoate synthase from limestone soil through metagenomic approach
    Tai YT, Foong CP, Najimudin N, Sudesh K
    J Biosci Bioeng, 2016 Apr;121(4):355-64.
    PMID: 26467694 DOI: 10.1016/j.jbiosc.2015.08.008
    PHA synthase (PhaC) is the key enzyme in the production of biodegradable plastics known as polyhydroxyalkanoate (PHA). Nevertheless, most of these enzymes are isolated from cultivable bacteria using traditional isolation method. Most of the microorganisms found in nature could not be successfully cultivated due to the lack of knowledge on their growth conditions. In this study, a culture-independent approach was applied. The presence of phaC genes in limestone soil was screened using primers targeting the class I and II PHA synthases. Based on the partial gene sequences, a total of 19 gene clusters have been identified and 7 clones were selected for full length amplification through genome walking. The complete phaC gene sequence of one of the clones (SC8) was obtained and it revealed 81% nucleotide identity to the PHA synthase gene of Chromobacterium violaceum ATCC 12472. This gene obtained from uncultured bacterium was successfully cloned and expressed in a Cupriavidus necator PHB(-)4 PHA-negative mutant resulting in the accumulation of significant amount of PHA. The PHA synthase activity of this transformant was 64 ± 12 U/g proteins. This paper presents a pioneering study on the discovery of phaC in a limestone area using metagenomic approach. Through this study, a new functional phaC was discovered from uncultured bacterium. Phylogenetic classification for all the phaCs isolated from this study has revealed that limestone hill harbors a great diversity of PhaCs with activities that have not yet been investigated.
    Matched MeSH terms: DNA Primers
  19. Fulltext Cross-amplification of microsatellite markers across agarwood-producing species of the Aquilarieae tribe (Thymelaeaceae)
    Pern YC, Lee SY, Ng WL, Mohamed R
    3 Biotech, 2020 Mar;10(3):103.
    PMID: 32099744 DOI: 10.1007/s13205-020-2072-2
    Tree species in the Aquilarieae tribe of the Thymelaeaceae family produce agarwood, a natural product highly valued for its fragrance, but the species are under threat due to indiscriminate harvesting. For conservation of these species, molecular techniques such as DNA profiling have been used. In this study, we assessed cross-amplification of microsatellite markers, initially developed for three Aquilaria species (A.crassna, A.malaccensis, and A.sinensis), on ten other agarwood-producing species, including members of Aquilaria (A.beccariana, A.hirta, A.microcarpa, A.rostrata, A.rugosa, A.subintegra, and A.yunnanensis) and Gyrinops (G.caudata, G.versteegii, and G.walla), both from the Aquilarieae tribe. Primers for 18 out of the 30 microsatellite markers successfully amplified bands of expected sizes in 1 sample each of at least 10 species. These were further used to genotype 74 individuals representing all the 13 studied species, yielding 13 cross-amplifiable markers, of which only 1 being polymorphic across all species. At each locus, the number of alleles ranged from 7 to 23, indicating a rather high variability. Four markers had relatively high species discrimination power. Our results demonstrated that genetic fingerprinting can be an effective tool in helping to manage agarwood genetic resources by potentially supporting the chain-of-custody of agarwood and its products in the market.
    Matched MeSH terms: DNA Primers
  20. Fulltext Psychrophilic lipase from arctic bacterium
    Zakiah Ramle, Rashidah Abdul Rahim
    Trop Life Sci Res, 2016;27(11):151-157.
    MyJurnal
    A lipase producer psychrophilic microorganism isolated from Arctic sample was
    studied. The genomic DNA of the isolate was extracted using modified CTAB method.
    Identification of the isolate by morphological and 16S rRNA sequence analysis revealed
    that the isolate is closely related to Arthrobacter gangotriensis (97% similarity).
    A. gangotriensis was determined as positive lipase producer based on the plate screening
    using specific and sensitive plate assay of Rhodamine B. The PCR result using
    Arthrobacter sp.’s full lipase gene sequence as the template primers emphasised a
    possible lipase gene at 900 bp band size. The gene is further cloned in a suitable vector
    system for expression of lipase.
    Matched MeSH terms: DNA Primers
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