猿猴免疫缺陷病毒的酶活性逆转录酶在细菌中的表达：对核苷酸类似物抑制剂的敏感性

Expression of enzymatically active reverse transcriptase of simian immunodeficiency virus in bacteria: sensitivity to nucleotide analogue inhibitors.

作者信息

Prasad V R, Myrick K, Haseltine W, Goff S P

机构信息

Department of Biochemistry, Columbia University, College of Physicians and Surgeons, New York, New York.

出版信息

Virology. 1990 Dec;179(2):896-900. doi: 10.1016/0042-6822(90)90164-m.

DOI:10.1016/0042-6822(90)90164-m

PMID:1700544

Abstract

A fragment of the SIVmac251 pol gene was expressed in Escherichia coli as a trpE fusion protein. Analysis of extracts from bacteria containing this expression plasmid revealed the presence of a reverse transcriptase activity dependent on Mg2+ as divalent cation and active on both poly(rA).oligo(dT) and poly(rC.oligo(dG) templates. In comparative studies, the SIV and HIV-1 reverse transcriptases expressed in bacteria displayed very similar high sensitivities to the chain terminator inhibitors AZTTP and ddTTP. The reverse transcriptase of Moloney murine leukemia virus and the DNA polymerase of E. coli were both more resistant to ddTTP, and the E. coli enzyme was significantly more resistant to AZTTP.

摘要

SIVmac251 pol基因的一个片段在大肠杆菌中作为trpE融合蛋白表达。对含有该表达质粒的细菌提取物的分析表明，存在一种依赖Mg2+作为二价阳离子的逆转录酶活性，并且对聚（rA）·寡聚（dT）和聚（rC）·寡聚（dG）模板均有活性。在比较研究中，在细菌中表达的SIV和HIV-1逆转录酶对链终止抑制剂AZTTP和ddTTP表现出非常相似的高敏感性。莫洛尼鼠白血病病毒的逆转录酶和大肠杆菌的DNA聚合酶对ddTTP的抗性都更强，并且大肠杆菌的酶对AZTTP的抗性明显更强。