• Title/Summary/Keyword: Single gene

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Construction of a Temperature Controlled Expression Ve e tor in Saccharumy ces cerevisiae (Saccharomyces cerevisiae를 이용한 온도조절형 발현 Vector의 개발)

  • 최진옥;황용일
    • Microbiology and Biotechnology Letters
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    • v.21 no.3
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    • pp.214-220
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    • 1993
  • The mating type a of yeast, Saccharomyces cerevisiae mutant with hmla2-102 and sir3-8ts was changed to type alpha by changing the growth temperature from 25C to 35C. A temperature-sensitive expression vector system was constructed using mating factor alpha1 (Mfalpha1) gene encoding alpha factor which is expressed in the type alpha cells. Vectors with different copy numbers were constructed by joining the promoter and pre or prepro-secretion single sequence of Mfalpha1 to promoterless PHO5' gene as a reporter gene.

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Single-minded 1 Gene Mapping and Its Variants Association with Growth, Carcass Composition and Meat Quality Traits in the Pig

  • Zhao, X.F.;Xu, N.Y.;Chen, Z.;Wang, Q.;Guo, X.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.7
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    • pp.941-946
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    • 2008
  • Single-minded 1 gene (SIM1) is a homolog of Drosophila SIM1 gene which plays a key role in the midline cell lineage of the central nervous system and is implicated in the regulation of feeding behavior and obesity in the human and mouse. In this study, porcine SIM1 gene was firstly mapped to chromosome 1p13 using radiation hybrid (RH) mapping and two polymorphisms were detected at position 607 (A/G) in SIM1 intron7 and position 780 (C/T) in SIM1 exon8. The last substitution was genotyped in a 364 F2 animal-population and an association analysis of these genotypes was performed with growth, carcass and meat quality traits by the statistical animal model. The results showed the significant influence of the SIM1 genotype on growth (p<0.05): live weight at birth, later period of growth and average daily gain; and effects on carcass composition (p<0.05): weight of head and buck kneed foreleg, backfat depth, loin eye area, carcass leaf fat and ham fat weights; and traits related to intramuscular fat content (p<0.05).

Genetic Variation in PDCD6 and Susceptibility to Lung Cancer

  • He, Yan-Qi;Zhou, Bin;Shi, Shao-Qing;Zhang, Lin;Li, Wei-Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4689-4693
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    • 2012
  • Lung cancer is the most common type of cancer and one of the leading causes of death in the world. Genetic factors play an important role in its development. PDCD6, the encoding gene for programmed cell death protein 6, may function as a tumor suppressor gene. Non-small cell lung cancer (NSCLC) contributes about 80% to newly histologically diagnosed lung cancer patients. To explore the relationship between PDCD6 and NSCLC, we examined two single nucleotide polymorphisms(rs3756712 G/T andrs4957014 G/T, both in the intron region) of the PDCD6gene.A hospital-based case-control study was carried out including 302 unrelated NSCLC patients and 306 healthy unrelated subjects. Significantly increased NSCLC risk was found to be associated with the T allele of rs4957014 (P=0.027, OR=0.760, 95%CI=0.596-0.970). The genotype and allele frequencies of rs3756712 did not shown any significant difference between NSCLC group and controls (P=0.327, OR=0.879, 95%CI=0.679-1.137). In conclusion, we firstly demonstrated the association between the PDCD6 gene and risk of NSCLC in a Chinese Han population.

Cloning and Expression in E. coli of the HOPDA Hydrolase Gene from Pseudomonas sp. P20

  • Lim, Jong Chul;Chae, Jong Chan;Kim Youngsoo;Kim, Hyong Bai;Kim Chi Kyung
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.349-354
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    • 1996
  • Pseudomonas sp. P20 is a natural isolate which is capable of degrading biphenyl and 4-chlorobiphenyl. From a clone of pCK1022 harboring pcbCD genes of Pseudomonas sp P20, a pcbD gene encoding 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoic acid (HOPDA) hydrolase was subcloned in Escherichia coli XL-1-Blue by using pBluescript SK(+) vector. The 2.8-kb HindII fragment harboring the pcbD gene cloned in pCK 1024 had a single site for each of XhoI, SalI, BstXI, and XbaI restriction enzymes. Escherichia coli CK1024 had a single site for each of XhoI, SalI, BstXI, and XbaI restriction enzymes. Escherichia coli CK1024 carrying pCK0124 degraded HOPDA to benzoate and 2-hydroxypenta-2, 4-dienoate by HOPDA hydrolase encoded by pcbD gene as effectively as E coli CK 1022 HARBORING pcbCD genes.

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Characterization of the BolA Homolog IbaG: A New Gene Involved in Acid Resistance

  • Guinote, Ines Batista;Moreira, Ricardo Neves;Freire, Patrick;Arraiano, Cecilia Maria
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.484-493
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    • 2012
  • BolA protein homologs are widely distributed in nature. In this report, we have studied for the first time YrbA, the only BolA homolog present in Escherichia coli, which we have renamed ibaG. We have constructed single and multiple ibaG mutants, and overexpressed ibaG in wild-type strains, in order to characterize this gene. The ibaG phenotypes are different from the bolA-associated round morphologies or growth profiles. Interestingly, ibaG and bolA single-and double-deletion mutants grow faster and have higher viabilities in rich media, whereas the overexpressed strains are significantly growth impaired. However, the mutant strains have lower viabilities than the wild type in the late stationary phase, indicating that both bolA and ibaG are important for survival in difficult growth conditions. bolA, as a transcription factor, binds to some promoters, but ibaG does not interact with the same DNA regions. We have determined that ibaG is transcribed in an operon with the murA gene, involved in the synthesis of peptidoglycan precursors. ibaG was also seen to change its mRNA expression pattern in response to acidic stress. ibaG may thus represent a new gene involved in cell resistance against acid stress.

Molecular Cloning of a Pepper Gene that Is Homologous to SELF-PRUNING

  • Kim, Dong Hwan;Han, Myeong Suk;Cho, Hyun Wooh;Jo, Yeong Deuk;Cho, Myeong Cheoul;Kim, Byung-Dong
    • Molecules and Cells
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    • v.22 no.1
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    • pp.89-96
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    • 2006
  • "Determinate" and "indeterminate" inflorescences in plants are controlled by a single recessive gene, for example, SELF-PRUNING (SP) in Solanum lycopersicum, TERMINAL FLOWER1 in Arabidopsis, CENTRORADIALIS in Antirrhinum, and CENTRORADIALIS-like gene in tobacco. Pepper (Capsicum annuum L.) is an indeterminate species in which shoots grow indefinitely. In this study, we cloned and characterized the pepper SP-like gene (CaSP). RT-PCR revealed that the CaSP transcript accumulates to higher levels in floral buds than in other organs. Comparison of genomic DNA and cDNA sequences from indeterminate and determinate pepper plants revealed the insertion of a single base in the first exon of CaSP in the determinate pepper plants. CaSP is annotated in linkage group 8 (chromosome 6) of the SNU2 pepper genetic map and showed similar synteny to SP in tomato. Transgenic tobacco plants overexpressing CaSP displayed late-flowering phenotypes similar to the phenotypes caused by overexpression of CaSP orthologs in other plants. Collectively, these results suggest that pepper CaSP is an ortholog of SP in tomato.

Direct Intramuscular Gene Transfer of Naked DNA Expressing Human Vascular Endothelial Growth Factor (pCK-VEGF) Enhances Collateral Growth in a Rabbit Ischemic Hind Limb Model (토끼 허혈성 하지 모델에서 VEGF 발현 Naked DNA 벡터인 pCK-VEGF의 근육내 투여가 측부혈관형성에 미치는 영향)

  • 채제건;전현순;박은진;김종묵;김덕경;김선영
    • YAKHAK HOEJI
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    • v.45 no.1
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    • pp.108-115
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    • 2001
  • We have recently reported the development of a high efficiency expression vector, pCK, which can drive a high level of gene expression in mouse skeletal muscle. In this study, we tested the therapeutic potential of pCK expressing human VEGF165, pCK-VEGF in the rabbit ischemic hind limb model. To determine the optimal dose of plasmid DNA, various concentrations of pCK-CAT were injected into the muscle of a rabbit hind limb and the levels of CAT activity were determined. It was found that the expression level of the exogenously added gene became stable between 250 and 1,000 $\mu$g. Based on this result, we tested whether intramuscular transfer of 500$\mu$g of pCK-VEGF could actually modulate collateral vessel development in a rabbit ischemic hind limb model. It was found that relative to the control group injected with the pCK lacking the VEGF sequence, single intramuscular doses (500$\mu$g) of pCK-VEGF produced statistically significant augmentation of collateral vessels as determined by the angiographic vessel count, maximal blood flow by Doppler flowmeter and the number of capillaries by histology. These results suggest that a single 500$\mu$g-delivery of pCK-VEGF is potent enough to induce sufficient angiogenic activity and achieve therapeutic benefit on this rabbit model.

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Single Somatic Embryogenesis from Transformant with Proteinase II Gene in Panax ginseng C.A. Meyer

  • Yang, Deok-Chun;Kim, Se-Young;Rho, Yeong-Deok;Kim, Moo-Sung
    • Plant Resources
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    • v.6 no.3
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    • pp.205-210
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    • 2003
  • Ginseng(Panax ginseng C.A. Meyer) is a perennial herbaceous plant which grows very slowly. It takes about 3 to 4 years from seeding to collecting the ripe seeds and the ginseng propagation is very difficult. and so, it is very difficult to breed ginseng plant. Ginseng tissue culture was started from at 1960, and ginseng commercial product by in vitro callus culture was saled, however upto now, regenerants were not planted to soil normally. Recently, plant genetic engineering to produce transgenic plants by introducing useful genes has been advanced greatly. In a present paper, transformation of ginseng plants was achieved by co-cultivation with Agrobacterium harboring the binary vector coding Proteinase-II gene, which confer resistant or tolerant to insect pests, The binary vector for transformation was constructed with disarmed Ti-plasmid and with double 35S promoter. The NPT II gene and introduced genes of the transgenic ginseng plants were successfully identified by the PCR. Especially the transgenic ginseng plants were regenerated using new techniques such as repetitive single somatic embryogenesis.

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Identification of Hanwoo and Holstein meat using MGB probe based real-time PCR associated with single nucleotide polymorphism (SNP) in Melanocortin 1 receptor (MC1R) gene (소 모색관련 MC1R 유전자의 SNP와 관련한 MGB probe에 기초한 real-time PCR을 이용한 한우육과 Holstein육의 판별)

  • Park, Sung-Do;Kim, Tae-Jung;Lee, Jae-Il
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.25-28
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    • 2005
  • The melanocortin 1 receptor (MC1R) plays an important role in regulation of melanin pigment synthesis within mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat color variations within several mammalian species including cattle. To develope a rapid and accurate method for the identification of Hanwoo meat, we performed a single nucleotide polymorphism (SNP) analysis in Melanocortin 1 receptor (MC1R) gene using TaqMan$^{(R)}$ MGB probe-based real-time PCR. Two specific probes (one for Hanwoo and the other for Holstein and Black angus) were designed. At the 5' end of 2 TaqMan$^{(R)}$ MGB probes, 6-carboxyfluorescein (FAM) was labeled for Hanwoo, and VIC for Holstein and Black angus. As a result, Hanwoo samples showed FAM-positive signal only, whereas other samples showed VIC-positive. This result suggests that the TaqMan$^{(R)}$ MGB probe based real-time PCR technique would be very accurate, easy and reproducible method to discriminate between Hanwoo meat and Holstein/Black angus meat.