目的克隆中国人谷胱甘肽过氧化物酶(GPx1)的cDNA。方法用逆转录聚合酶链反应(RT-PCR)，以中国人胎盘组织总RNA为模板，扩增中国人谷胱甘肽过氧化物酶(GPx1)的cDNA，进行序列分析。计算机分析其二级结构并比较已发现的人GPx家族的5种GPx氨基酸序列。结果从中国人胎盘组织中克隆的GPx1基因，与国外文献报道相比，只有1个氨基酸残基发生变异，即leu22变为Val22，该变化不影响GPx1活性中心的结构；人GPx的硒结合区及活性中心区域的氨基酸序列是高度保守的。结论首次克隆了中国人体特异的胞质内GPx1基因，为其生物学活性及结构与功能的关系的研究创造了条件。
Abstract
OBJECTIVE To clone the cDNA of human glutathione peroxidase 1(GPx1).METHODS The total RNA was isolated from normal Chinese human placenta and was amplified as the template by RT-PCR.It was inserted into pSK plasmid to analyze its sequence;the sequences of 5 known GPx gene were compared. RESULTS The GPx1 cDNA from placenta had 1 base pare and 1 amino acid residue (Leu22mutated to Val22),different from that of reported.But it didn't alter the secondly structure of GPx selenocysteint residue and the active sites were highly conserved in 5 GPxs. CONCLUSION The human GPx1 gene of Chinese was cloned for the first time.
关键词
谷胱甘肽氧化物酶 /
基因克隆 /
序列分析 /
GPx家族 /
超氧化物歧化酶
{{custom_keyword}} /
Key words
glutathione peroxidase /
gene cloning /
sequence analysis
{{custom_keyword}} /
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Takahashi K,Avissar N,Whitin J, et al. Purification and characerization of human glutsthione peroxidase:A selenoglycoprotein distinct from the known cellular enzyme[J]. Arch Biochem Biophys, 1987,256:677～686.
[2] Sukenaga Y,Ishida K,Takeda T, et al. cDNA sequence coding for human gmutathione peroxidase[J]. Nucleic Acids Res, 1987,15(17):7178.
[3] Chu FF,Doroshow JH,Esworthy RS.Expression ,characterization,and tissue distribution of a new cellular seleniumdependent glutathione peroxidase,GSHPxGI[J]. J Biol Chem, 1993,268(4):2571～2576.
[4] Takahashi K,Akasaka M,Yamamoto Y, et al. Primary structure of human plasma glutathione peroxidase deduced from cDNA sequences[J]. J Biochem, 1990,108(2):145～148.
[5] Kelner MJ,Montoya MA.Structural organization of the human seleniumdependent phospholipid hydroperoxide glutathione peroxidase gene (GPx4):chromosomal localization to 19p13.3[J]. Biochem Biophys Res Commun, 1998,249(1):53～55.
[6] Hall L,Williams K,Perry ACF, et al. The majority of human glutathione peoxidase type 5(GPx5) transcripts are incorrectly spliced:implications for the role of GPx5 in the male reproductive tract[J]. Biochem J, 1998,333:5～9.
[7] Bravard A,Petridis F,Luccioni C.Modulation of antioxidant enzymes p21 WAFI and p53 expression during proliferation and differentiation of human melanoma cell lines[J]. Free Radic Biol Med, 1999,26(7～8):1027～1033.
[8] 孙娟，陈瑗，周玫.GPx对活性氧诱发细胞程序性死亡的影响[J].生物化学与生物物理进展,1998,25(3):259～261.
[9] Tan M,Li S,Swaroop M, et al. Transcriptional activation of the human glutathione peroxidase promoter by p53[J]. J Biol Chem,1999,274(17):12061～12066.
[10] Sambrook J,Fritsch EF,Maniatist T. Molecular cloning:A Laboratotry Manual. 2nd ed[M].New York:Cold Spring Harbor Laboratory Press,1989.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
