人类cyclophilin A基因cDNA的克隆和序列测定cDNA Cloning and Sequencing of Human Cyclophilin A Gene
任志娟,王季石,肖芸,张维,陈剑,孙等军,张旭,吴镇宇,方琴
REN Zhi-juan ; WANG Ji-shi ; Xiao Yun ; ZHANG Wei; CHEN Jian ; SUN Deng-jun; ZHANG Xu ; WU Zhen-yu; FANG Qin
;
摘要(Abstract):
目的:克隆人类环孢霉素A受体亚型cyclophilinA(CypA)基因,对该基因进行序列测定。方法:利用EST重叠序列拼排技术,设计特异性CypA引物,以人外周血白细胞总RNA为模板,进行RT-PCR扩增,纯化PCR产物,装pGEM-T载体,进行DNA序列测定。结果:成功克隆了人类Cyp A基因cDNA序列的开放阅读框,经DNA序列测定证实序列正确。结论:通过克隆人CypA基因,并克隆人pGEM-T载体,为下一步的基因表达和功能研究奠定了实验基础。
Objective: To clone and analyze cDNA sequence of human cyclophilin A gene. Methods: The EST (expressed sequence tag) sequence alognment method, RT-PCR,DNA sequencing, and a computer biology software were used for the cDNA cloning and sequence analysis. Results: The full length of cDNA of human cyclophilin A gene was successfully cloned and sequenced. Conclusion: This study provides a foundation for further gene expression experiment and the next step of function research.
关键词(KeyWords):
基因组;人;克隆;分子;受体;环孢霉素
genome, human; cloning, molecular; receptor, cyclosporins
基金项目(Foundation): 贵州省省长基金(S2000-15)
作者(Authors):
任志娟,王季石,肖芸,张维,陈剑,孙等军,张旭,吴镇宇,方琴
REN Zhi-juan ; WANG Ji-shi ; Xiao Yun ; ZHANG Wei; CHEN Jian ; SUN Deng-jun; ZHANG Xu ; WU Zhen-yu; FANG Qin
;
DOI: 10.19367/j.cnki.1000-2707.2003.02.008
参考文献(References):
- [1] Santosh A, Helekar, Patrick J, et al. Peptidyl prolyl cistrans isomerase activity of cyclophilin A in functional homo-oligomeric receptor expression [J]. Pnas, 1997,94:5 432-5 437.
- [2] Sherry B, Zybarth G, Alfao M, et al. Role of cyclophilin A in the uptake of HIV-1 by macrophages and T lymphocytes [J]. Pnas, 1998,95: 1 758-1 763
- [3] Brazin KN, Mallis RJ, Fulton DB, et al. Regulation of the tyrosine kinase by the peptidyl-prolyl isomerase cyclophilin A [J]. Pnas, 2002, 99: 1 899-1 904.
- [4] Braaten D, Luban J. Cyclophilin A regulates HIV-1 infectivity, as demonstrated by gene targeting in human Tcells [J]. EMBO J,2001, 20: 1 300-1 309
- [5] Fitzon T, Leschonsky B, Bieler K, et al. Proline residues in the HIV-1 NH2-teminal capsid domain: structure determinants for proper core assumbly and subsequent steps of early replication [J]. Virology, 2000,268:294-307.
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- 任志娟
- 王季石
- 肖芸
- 张维
- 陈剑
- 孙等军
- 张旭
- 吴镇宇
- 方琴
REN Zhi-juan - WANG Ji-shi
- Xiao Yun
- ZHANG Wei
- CHEN Jian
- SUN Deng-jun
- ZHANG Xu
- WU Zhen-yu
- FANG Qin
- 任志娟
- 王季石
- 肖芸
- 张维
- 陈剑
- 孙等军
- 张旭
- 吴镇宇
- 方琴
REN Zhi-juan - WANG Ji-shi
- Xiao Yun
- ZHANG Wei
- CHEN Jian
- SUN Deng-jun
- ZHANG Xu
- WU Zhen-yu
- FANG Qin