• Title/Summary/Keyword: J gene

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Primary Structure of the Human VkII Regions Elicited by Haemophilus influenzae Type b Polysaccharide Vaccines; The J Gene Usage Is Restricted in Child Antibodies Using the A2 Gene

  • Yu, Kang-Yeol;Kim, Jin-Ho;Chung, Gook-Hyun
    • BMB Reports
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    • v.33 no.3
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    • pp.249-255
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    • 2000
  • The Haemophilus influenzae type b (Hib) has been a major cause of bacterial meningitis in children who are less than two years old. The variable (V) region repertoire of adult Caucasian antibodies (Abs) to Hib polysaccharide (PS) has been characterized well. The majority of adult antibodies against Hib uses VL that is derived from the Vk gene A2 and have arginine at the N region. In order to explore the possibility those antibody responses to Hib-PS is variable in various age groups, we examined the VL regions of the antibodies to Hib-PS in Korean adults and children. We immunized Korean adults (n = 8) and children (n = 39) with Hib tetanus conjugated vaccines, isolated RNAs from the peripheral lymphocytes, and amplified the A2-derived VL regions by RT-PCR. The PCR products were subcloned and sequenced. Forty-seven out of 54 independent clones from children used the $J{\kappa}2$, or $J{\kappa}3$ gene in preference. The adults, however, used all of the $J{\kappa}$ genes evenly. With respect to the amino acid sequences of variable regions, adult $A2-J{\kappa}$ recombinants have a germline sequence. But, the 76th codon (AGC) of child $A2-J{\kappa}2$ recombinants was substituted with CGC (arginine) in most cases (88 %) and 77 percent of child clones using the $A2-J{\kappa}3$ genes have isoleucine-109 at the junction of $J{\kappa}-C{\kappa}$ instead of threonine that is found in a germline sequence. These results suggest that the mechanism of antibody production in young children is different from that of adults.

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Expression of B Cell Activating Factor Pathway Genes in Mouse Mammary Gland

  • Choi, S.;Jung, D.J.;Bong, J.J.;Baik, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.2
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    • pp.153-159
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    • 2007
  • In our previous study, overexpression of extracellular proteinase inhibitor (Expi) gene accelerated apoptosis of mammary epithelial cells, and induced expression of B cell activating factor (BAFF) gene. In this study, we found induction of BAFF-receptor (BAFF-R) gene expression in the Expi-transfected cells. A proliferation-inducing ligand (APRIL) gene is another TNF family member and the closest known relative of BAFF. We found induction of APRIL gene expression in the Expi-overexpressed apoptotic cells. NF-${\kappa}$B gene was also induced in the Expi-overexpressed cells. Expression patterns of BAFF and APRIL pathway-related genes were examined in in vivo mouse mammary gland at various reproductive stages. Expression levels of BAFF gene were very low at early pregnancy, increased from mid-pregnancy, and peaked at lactation, and thereafter decreased at involution stages of mammary gland. Expression of BAFF-R gene was highly induced in involution stages compared to lactation stages. Thus, expression patterns of BAFF-R gene were correlated to apoptotic status of mammary gland: active apoptosis of mammary epithelial cells occurs at involution stage of mammary gland. Expression levels of NF-${\kappa}$B gene were higher in involution stages compared to lactation stages. We analyzed mRNA levels of bcl-2 family genes from different stages of mammary development. Bcl-2 gene expression was relatively constant during lactation and involution stages. There was a slight increase in bcl-xL gene expression in involution stages compared to lactation state. Bax gene expression was highly induced in involution stage. Our results suggest that signaling pathways activated by both BAFF and ARRIL in mammary gland point towards NF-${\kappa}$B activation which causes upregulation of bax.

A Macrolide-Lincosamide-Streptogramin B Resistance Determinant Gene (ermJ) Cloned from B, anthracis 590

  • Kim, Hee-Sun;Choi, Eung-Chil;Kim, Byong-Kak;Park, Young-In
    • Archives of Pharmacal Research
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    • v.15 no.1
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    • pp.58-61
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    • 1992
  • Bacillus anthracis 590 having an inducibla resistance determinant to MLS antibiotics was isolated from a soli sample in Korea. The resistance gene (ernJ) was cloned by Southern blotting of chromosomal DNA fragment digested by various restriction enzymes and coloy hybridization method and the cloned plasmid was named as pBA423. The size of inserted DNA fragment of pBS42 vector was about 2.9 kb and the DNA sequence of the subcloned fragment (Hinc II-Hinc II, 1.4kb) WAS determined. The DNA sequence of ernJ was composed of 357 bp for leader region and 861 bp for the structural gene. Because the leader sequence of ernJ was homologous to that of ermK, the expression of ernJ is also thought to be controlled by a transcriptionl attenuation mechanism.

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A Single Nucleotide Polymorphism in LOC534614 as an Unknown Gene Associated with Body Weight and Cold Carcass Weight in Hanwoo (Korean Cattle)

  • Lee, Y.S.;Oh, D.Y.;Kim, J.J.;Lee, J.H.;Park, H.S.;Yeo, J.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.12
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    • pp.1543-1551
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    • 2010
  • A major aim of cattle genome research is to identify candidate genes associated with meat quantity and quality through QTL analysis for application in the livestock industry. Therefore, this study focused on discovery of useful SNPs within the LOC534614 gene, containing 12273_165 SNP which is located on the same site as the QTL on chromosome 6, and evaluation of the association between SNP and body weight and cold carcass weight in Hanwoo (Korean cattle) As a result of a BLAST search of the NCBI web site, we discovered that the mRNA sequence of the LOC534614 gene was similar to that of the coiled-coil domain containing 158 (CCDC158) for dog and human. According to the direct DNA sequence from the CCDC158 gene, we identified 19 polymorphic SNPs within exons and their flanking regions. Among them, 17 polymorphic SNPs were selected for genotyping in Hanwoo (n = 476) and seventeen marker haplotypes containing 12273_165 SNP (frequency >0.1) were identified. As a result of the association between 17 polymorphic SNPs and Hanwoo (n = 476), g.8778G>A SNP in exon 6 was found to be a non-synonymous SNP, and was significantly associated with body weight and cold carcass weight (p<0.05). We discovered 19 polymorphic SNPs in the CCDC158 gene on the QTL region of BTA 6 in Hanwoo and identified that the g.8778G>A SNP was significantly associated with body weight and cold carcass weight (p<0.05), which causes an amino acid variation from valine to methionine. Furthermore, statistical analysis demonstrated that the CCDC158 gene is strongly associated with body weight and cold carcass weight in Hanwoo. In this regard, the g.8778G>A SNP in the CCDC158 gene can be useful as a positional candidate for body weight and cold carcass weight for marker-assisted selection in Hanwoo.

Expression of Green Fluorescent Protein(GFP) Gene in Rabbit Embryos (토끼 수정란에서 Green Fluorescent Protein 유전자의 발현)

  • Kang, T. Y.;Yin, X. J.;Chae, Y. J.;Lee, H.;Lee, H. J
    • Journal of Embryo Transfer
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    • v.14 no.1
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    • pp.1-8
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    • 1999
  • The efficiency of transgenic livestock animal production may be improved by early selection of transgenci preimplantation embryos. To examine the possibility of GFP gene as a non-invasive marker for the early screening of transgenic embryo, the GFP gene was microinjected into rabbit zygotes and the later stages of preimplantation embryos were examined for the expression of GFP. The presence of injected DNA was detected by PCR analysis and the expression of GFP was detected by observing green fluorescence in embryos under a fluorescent microscope. Out of 108 GFP gene-injected rabbit zygotes, seventy three(67.6%) were fluorescence-positive. When 11 fluroresecence-positive blastocysts were analyzed for the presence of GFP gene by PCR, 6(54.5%) were positive, and all of the 8 flrouescence-negative blastocysts were also negative by PCR. The results indicate that the screening of transgene in rabbit embryos by PCR analysis and GFP detection could be a promising method for the preselection of transgenic embryos.

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DNA-dependent Protein Kinase Mediates V(D)J Recombination via RAG2 Phosphorylation

  • Hah, Young-Sool;Lee, Jung-Hwa;Kim, Deok-Ryong
    • BMB Reports
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    • v.40 no.3
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    • pp.432-438
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    • 2007
  • V(D)J recombination, a site-specific gene rearrangement process occurring during the lymphocyte development, begins with DNA double strand breaks by two recombination activating gene products (RAG1/2) and finishes with the repair process by several proteins including DNA-dependent protein kinase (DNA-PK). In this report, we found that RAG2 was specifically phosphorylated by DNA-PK at the $365^{th}$ serine residue, and this phosphorylated RAG2 affected the V(D)J recombination activity in cells in the GFP expression-based assay. While the V(D)J recombination activity between wild-type RAG2 and mutant S365A RAG2 in the assay using a signal joint substrate was undistinguishable in DNA-PK deficient cells (M059J), the activity with wild-type RAG2 was largely increased in DNA-PK proficient cells (M059K) in comparison with mutant RAG2, suggesting that RAG2 phosphorylation by DNA-PK plays a crucial role in the signal joint formation during V(D)J recombination.