• Title/Summary/Keyword: J gene

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Cloning and Sequencing of Heterozygous PSS Gene in Pigs (Cloning을 이용한 PSS Hetero 돼지에서의 염기 서열 분석)

  • Yoo J. Y.;Kim G. W.;Lee J. W.;Kim Y. B.;Lee J. Y.;Lee D. H.;Lee H. J.;Yoon J. M.;Park H. Y.
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.15-18
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    • 2005
  • This study was carried out to analyze the structure of the gene related to porcine stress syndrome (PSS) through cloning from PSS heterozygous pigs and to examine hereditary type associated with PSS. The results obtained from the study were summarized as follows: Amino acid arginine and cysteine exist in dominant gene, N and harmfully recessive gene, n, respectively. It was confirmed that such genes exists as alleles in the sixth chromosome related to PSS. These allelic genes might be inherited according to Mendelian law.

Variation of fibrinolytic enzyme activity produced Bacillus subtilis by gene cloning (유전자 cloning에 의한 Bacillus subtilis의 fibrinolytic enzyme 활성 변화)

  • 이홍석;유천권;이철수;강상모
    • Microbiology and Biotechnology Letters
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    • v.28 no.1
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    • pp.14-20
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    • 2000
  • The transformation of Bacillus subtilis K-54 and J-10 was carried out with constructed vectors containing structure and enhancer genes of aprN and prtR, to increase their fibrinolytic enzyme activity. Bands for the aprN and prtR genes were identified from B. subtilis J-10 by PCR that was carried out with the constructed primers for the genes. In addition, the gene fragments contained promoter site based on the results of analysing their nucleotide sequence. The two gene fragments, aprN and prtR, obtained by the PCR, were, then, inserted to vector such as T-vector and E.coli/Bacillus shuttle vector. The constructed vector were designated as pAPR2 (aprN), pENC2 (prtR) and pFLA1 (aprN and prtR), respectively. The constructed vector was used for transformation of the strains of B.subtilis J-10 and B. subtilis K-54 and the fribrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and the fibrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and pFLA1, resulted in the increase of fibrinolyitic enzyme activity in B. subtilis J-10 by 27.3% and 16%, respectively. However, the introduction of pENC2 to B. subtilis J-10 did not seem to induce increase of the enzyme activity. The strain of B.subtilis K-54 transformed with pENC2 showed an increased fibrinolytic activity by 5 folds compared with that of the original strain of B. subtilis K-54.

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Identification of Candidate SNP (Single Nucleotide Polymorphism) for Growth and Carcass Traits Related to QTL on Chromosome 6 in Hanwoo (Korean Cattle)

  • Lee, Y.S.;Lee, J.H.;Lee, J.Y.;Kim, J.J.;Park, H.S.;Yeo, J.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.12
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    • pp.1703-1709
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    • 2008
  • As genetic markers, single nucleotide polymorphisms (SNP) are very appropriate for the development of genetic tests for economic traits in livestock. Several microsatellite markers have been identified as useful markers for the genetic improvement of Hanwoo. Among those markers, ILSTS035 was recently mapped at a similar position with four SNPs (AH1_11, AH1_9, 31465_446, and 12273_165) in a linkage map of EST-based SNP in BAT6. Among the four SNPs, two SNPs (31465_446 and 12273_165) were analyzed using BLAST at the NCBI web site. The sequences including the 12273_165 SNP were identified at the intron region within the LOC534614 gene on the gene sequence map (Bos taurus NCBI Map view, build 3.1). The LOC534614 gene represents a protein similar to myosin heavy chain, fat skeletal muscle, embryonic isoform 1 in the dog, and myosin_1 (Myosin heavy chain D) in Macaca mulatta. In cattle, the myosin heavy chain was associated with muscle development. The phenotypic data for growth and carcass traits in the 415 animals were analyzed by the mixed ANCOVA (analysis of covariance) linear model using PROC GLM module in SAS v9.1. By the genotyping of Hanwoo individuals (n = 415) to evaluate the association of SNP with growth and carcass traits, it was shown that the 12273_165 SNP region within LOC534614 may be a candidate marker for growth. The results of the statistical analyses suggested that the genotype of the 12273_165 SNP significantly affected birth weight, weight of the cattle at 24 months of age, average daily gain and carcass cold weight (p<0.05). Consequently, the 12273_165 SNP polymorphisms at the LOC534614 gene may be associated with growth in Hanwoo, and functional validation of polymorphisms in LOC534614 should be performed in the future.

Effect of Pronuclear Injection with Human Growth Hormone Gene on Development and PCR-Screening in Rabbit Embryos (사람성장호르몬 유전자의 전핵내 미세주입이 토끼 수정란의 체외발달에 미치는 영향과 PCR검색)

  • Kang, T. Y.;Chae, Y. J.;Lee, H.;Lee, K. K.;Park, C. S.;Lee, H. J.
    • Journal of Embryo Transfer
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    • v.13 no.2
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    • pp.97-106
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    • 1998
  • The pronuclear injection of metallothionein-human growth hormone (MT-hGH) gene into rabbit zygotes was performed to establish in vitro developmental system and to detect the presence of the injected gene by nested PCR. Mature female New Zealand White rabbits were superovulated by eGG and hCG treatments. The rabbits were mated and the zygotes were collected from the oviducts 18-22 h after hCG injection by flushing with D-PBS. Two to three picoliters of MT-hGH gene was microinjected into male pronuclei. The foreign gene-injected zygotes were cultured in TCM-199 or RD mediurn containing 10% FCS with a monolayer of rabbit oviductal epithelial cefls in a 5% $CO_2$ incubator. The presence of injected DNA in rabbit embryos or blastomeres at different developmental stages .vas detected by a nested PCR analysis. The results are summarized as follows ; 1.The developmental rate of the MT-hGH gene-injected zygotes to blastocyst was significantly higher in TCM-199 medium (68.1%) than in RD medium (42.9%). 2.The gene injection into pronuclei at 18 or 22 hours post hCG treatment during pronuclear stage did not much affect on the in vitro development of the rabbit embryos. 3.The rate of gene-positive embryos detected by the nested PCR analysis was significantly decreased when they developed to blastocysts. The results indicate that the screening of transgene in rabbit embryos by nested PCR analysis could be a prornisible method for the preselection of transgenic embryos. Furthermore, the preselection of transgenic embryos would greatly reduce hoth the cost and effort of production of transgenic animals.

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Pyramiding transgenes for potato tuber moth resistance in potato

  • Meiyalaghan, Sathiyamoorthy;Pringle, Julie M.;Barrell, Philippa J.;Jacobs, Jeanne M.E.;Conner, Anthony J.
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.293-301
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    • 2010
  • The feasibility of two strategies for transgene pyramiding using Agrobacterium-mediated transformation was investigated to develop a transgenic potato (Solanum tuberosum L. cv. Iwa) with resistance to potato tuber moth (PTM) (Phthorimaea operculella (Zeller)). In the first approach, cry1Ac9 and cry9Aa2 genes were introduced simultaneously using a kanamycin (nptII) selectable marker gene. The second approach involved the sequential introduction (re-transformation) of a cry1Ac9 gene, using a hygromycin resistance (hpt) selectable marker gene, into an existing line transgenic for a cry9Aa2 gene and a kanamycin resistance (nptII) selectable marker gene. Multiplex polymerase chain reaction (PCR) confirmed the presence of the specific selectable marker gene and both cry genes in all regenerated lines. The relative steady-state level of the cry gene transcripts in leaves was quantified in all regenerated lines by real-time PCR analysis. Re-transformation proved to be a flexible approach to effectively pyramid genes for PTM resistance in potato, since it allowed the second gene to be added to a line that was previously identified as having a high level of resistance. Larval growth of PTM was significantly inhibited on excised greenhouse-grown leaves in all transgenic lines, although no lines expressing both cry genes exhibited any greater resistance to PTM larvae over that previously observed for the individual genes. It is anticipated that these lines will permit more durable resistance by delaying the opportunities for PTM adaptation to the individual cry genes.

Induction of Lysozyme Gene Expression During Involution of Mouse Mammary Gland

  • Lee, M.J.;Han, O.;Back, K.;Choi, Y.J.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.4
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    • pp.462-466
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    • 2001
  • To understand molecular mechanisms of mouse mammary gland involution, clones were isolated by differential screening of a cDNA library. Partial sequences of a clone showed 100% identity to cDNA sequences of mouse lysozyme P gene. Northern analysis was performed to examine expression levels of lysozyme mRNA in mammary gland at several physiological states. Expression of lysozyme gene was induced at involution day 5 compared with lactating stage. High levels of lysozyme mRNA were also detected at virgin tissues. Two types of separate genes, P and M lysozyme, have been known in mouse, and we found that both lysozyme P and M genes were expressed in mammary tissues by reverse transcriptase-polymerase chain reaction. The lysozyme enzyme activity determined by lysoplate assay was also higher in involuted mammary tissues compared with lactating tissues, showing a similar trend to its mRNA levels. Lysozyme is an antimicrobial protein and involved in host defense mechanism. The increase in lysozyme gene expression may help to prevent microbial infection during mammary gland involution at which stage the residual milk in the mammary gland provides good nutritional sources for microbial growth.