• Title/Summary/Keyword: DnaJ

Search Result 704, Processing Time 0.035 seconds

Identification of Heterodera glycines (Tylenchida; Heteroderidae) Using qPCR

  • Ko, Hyoung-Rai;Kang, Heonil;Park, Eun-Hyoung;Kim, Eun-Hwa;Lee, Jae-Kook
    • The Plant Pathology Journal
    • /
    • v.35 no.6
    • /
    • pp.654-661
    • /
    • 2019

  • The soybean cyst nematode, Heterodera glycines, is a major plant-parasitic nematode that has caused important economic losses to Korea's soybean production. Four species of cyst nematodes, H. schachtii, H. glycines, H. trifolii, and H. sojae, all belong to schachtii group are coexist in field soil in Korea. The rapid identification of the nematode is crucial for preventing crop damage and in decision making for controlling this nematode. This study aimed to develop a species-specific primer set for quantitative PCR (qPCR) assay of H. glycines. The specific primer set (HGF1 and HGR1) for H. glycines was designed based on the cytochrome c oxidase subunit I (COI) sequence of mitochondrial DNA. After optimization, it is possible to identify the H. glycines using a qPCR assay with DNA extracted from a single cyst and single second-stage juvenile (J2). The specificity was confirmed by the absence of SYBR fluorescent signals of three other Heterodera species. A serial dilution of DNA extracted from a single cyst was obtained for the sensitivity test. The result showed that the standard curve of the test had a highly significant linearity between DNA concentration and Ct value (R2 = 0.996, slope = -3.49) and that the detection limit concentration of DNA of the primer set was 10 pg of DNA per reaction. Our findings suggested that H. glycines could be distinguished from H. sojae and other Heterodera species when a qPCR assay is used with a specific primer set.

  • https://doi.org/10.5423/PPJ.OA.04.2019.0086 인용 PDF KSCI

Full Length cDNA, Genomic Organizations and Expression Profiles of the Porcine Proteasomal ATPases PSMC5 Gene

  • Wang, Y.F.;Yu, M.;Liu, B.;Fan, B.;Wang, H.;Zhu, M.J.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.17 no.7
    • /
    • pp.897-902
    • /
    • 2004

  • PSMC5 subunit, which belongs to the 26S proteasomal subunit family, plays an important role in the antigen presentation mediated by MHC class I molecular. Full-length cDNA of porcine PSMC5 was isolated using the in silico cloning and rapid amplification of cDNA ends (RACE). Amino acid was deduced and the primary structure was analyzed. Results revealed that the porcine PSMC5 gene shares the high degree of sequence similarity with its mammalian counterparts at both the nucleotide level and the amino acid level. The RT-PCR was performed to detect the porcine PSMC5 expression pattern in seven tissues and the result showed that high express level was observed in spleen, lung, marrow and liver while the low express level was in muscle. The full-length genomic DNA sequence of porcine PSMC5 gene was amplified by PCR and the genomic structure revealed that this gene was comprised by 12 exons and 11 introns. Best alignment of the cDNA and genomic exon DNA sequence presents 4 mismatches and this information potentially bears further study in gene polymorphisms.

  • https://doi.org/10.5713/ajas.2004.897 인용 PDF KSCI

Identification of a Sequence Containing Methylated Cytidine in Corynebacterium glutamicum and Brevibacterium flavum Using Bisulfite DNA Derivatization and Sequencing

  • Jang, Ki-Hyo;Chambers, Paul J.;Britz, Margaret L.
    • Journal of Microbiology and Biotechnology
    • /
    • v.11 no.5
    • /
    • pp.819-824
    • /
    • 2001

  • The principal DNA modification systems of the amino-acid-producing bacteria Corynebacterium glutamicum AS019, Brevibacterium flavum BF4, and B. lactofermentum BL1 was investigated using two approaches; digestion of plasmid DNA isolated from these species TseI and Fnu4HI, and sequence analysis of the putative methyltransferase target sites following the derivatization of DNA using metabisulfite treatment. The C. glutamicum and B. flavum strains showed similar digestion patterns to the two enzymes, indicating that the target for cytidine methyltransferase recognizes 5'-GCSGC-3'(where S is either G or C). Mapping the methylated cytidine sites by bisulfite derivatization, followed by PCR amplification and sequencing, was only possible when the protocol included an additional step eliminating any underivatized DNA after PCR amplification, thereby indicating that the derivatization was not $100\%$ efficient. This may have been due to the high G0C content of this genus. It was confirmed that C. glutamicum AS019 and B. flavum BF4 methylated the cytidine in the $Gm^5CCGC$ sequences, yet there were no similar patterns of methylation in B. lactofermentum, which was consistent with the distinctive degradation pattern seen for the above enzymes. These findings demonstrate the successful application of a modified bisulfite derivatization method with the Corynebacterium species for determining methylation patterns, and showed that different species in the geneus contain distinctive restriction and modification systems.

  • PDF