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在人巨细胞病毒AD169的BamHI J片段中检测IE反应元件。

Detection of an IE responsive element(s) in the BamHI J fragment of human cytomegalovirus AD169.

作者信息

Reifel-Miller A E, Lee C H

机构信息

Department of Pathology, Indiana University School of Medicine, Indianapolis 46202.

出版信息

Virology. 1990 Aug;177(2):496-504. doi: 10.1016/0042-6822(90)90514-r.

Abstract

The BamHI J fragment of human cytomegalovirus (HCMV) AD169 located at 0.815 to 0.855 map units in the unique short component of the genome was demonstrated to be responsive to the HCMV IE proteins by using a transient chloramphenicol acetyltransferase (cat) gene expression system. The BamHI J fragment was cloned into a cat gene expression plasmid and then cotransfected with a plasmid that expresses the immediate early (IE) genes of HCMV AD169 into the HCMV permissive cell line MRC-5. The results indicated that the BamHI J fragment enhanced cat gene expression 10-fold when the HCMV IE proteins were present. The BamHI J fragment was demonstrated to have properties of an inducible enhancer. In the presence of the HCMV IE proteins, it enhances cat gene expression when positioned in either orientation both upstream and downstream from the cat gene; it enhances transcription from the herpes simplex virus type 1 (HSV-1) thymidine kinase gene and the simian virus 40 (SV40) early gene promoters; and it requires a cis-positioned promoter for enhancer activity.

摘要

人巨细胞病毒(HCMV)AD169的BamHI J片段位于基因组独特短片段的0.815至0.855图单位处,通过使用瞬时氯霉素乙酰转移酶(cat)基因表达系统,证明其对HCMV IE蛋白有反应。将BamHI J片段克隆到cat基因表达质粒中,然后与表达HCMV AD169立即早期(IE)基因的质粒共转染到HCMV允许细胞系MRC-5中。结果表明,当存在HCMV IE蛋白时,BamHI J片段可使cat基因表达增强10倍。已证明BamHI J片段具有诱导型增强子的特性。在HCMV IE蛋白存在的情况下,当它以任何方向位于cat基因的上游和下游时,均可增强cat基因表达;它可增强单纯疱疹病毒1型(HSV-1)胸苷激酶基因和猿猴病毒40(SV40)早期基因启动子的转录;并且其增强子活性需要一个顺式定位的启动子。

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