RETROVIRUS LONG TERMINAL REPEATS ACTIVATE EXPRESSION OF CODING SEQUENCES FOR THE HERPES-SIMPLEX VIRUS THYMIDINE KINASE GENE
RETROVIRUS LONG TERMINAL REPEATS ACTIVATE EXPRESSION OF CODING SEQUENCES FOR THE HERPES-SIMPLEX VIRUS THYMIDINE KINASE GENE
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DOI：
10.1073/pnas.79.5.1573
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发表时间：
1982-01-01
期刊：
影响因子：
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通讯作者：
BERNSTEIN, A
中科院分区：
文献类型：
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作者：
JOYNER, A;YAMAMOTO, Y;BERNSTEIN, A
小鼠逆转录病毒的长末端重复序列（LTR）可以激活缺失功能启动子区的异源基因编码序列的表达。获得了同时含有Friend脾局灶形成病毒（SFFV） DNA克隆片段和单纯疱疹病毒（HSV）胸苷激酶（TK； ATP：胸苷5 '-磷酸转移酶，EC 2.7.1.21）基因（TK）的重组质粒克隆。通过在SFFV 5”LTR的下游插入200或1200个碱基对，构建含有或不含启动转录所需的5”序列的tk编码序列的克隆，确定LTR对tk表达的影响，并通过基因转移到tk -小鼠细胞和tk +转化子中tk酶活性的测定来检测这些克隆对tk表达的影响。SFFV 5 " LTR激活表达tk当这些序列编码序列插入200个碱基对下游和相同的取向LTR. tk放置1200个碱基对也没激活下游和相同的取向LTR或当tk插入站点在相反方向的LTR。SFFV 5”LTR不会干扰体内的表达时tk两侧同源5”启动子序列。这些观察结果对逆转录病毒肿瘤发生和动物细胞遗传学的意义进行了讨论。
The long terminal repeats (LTR) of a murine retrovirus can activate expression of heterologous gene coding sequences from which a functional promoter region was deleted. Recombinant plasmid clones were obtained that contained both cloned fragments of Friend spleen focus-forming virus (SFFV) DNA and the herpes simplex virus (HSV) thymidine kinase (TK; ATP:thymidine 5''-phosphotransferase, EC 2.7.1.21) gene (tk). The effects of the LTR on tk expression were determined by constructing clones containing tk coding sequences with or without 5'' sequences necessary for the initiation of transcription, inserted either 200 or 1200 base pairs downstream from the SFFV 5'' LTR. The expression of the HSV TK protein by these clones was tested by gene transfer of the clones into TK- mouse cells and assay of TK enzyme activity in TK+ transformants. The SFFV 5'' LTR activates expression of tk coding sequences when these sequences are inserted 200 base pairs downstream from and in the same orientation as the LTR. tk is not activated when placed 1200 base pairs downstream from and in the same orientation as the LTR or when tk is inserted in either site in the opposite orientation as the LTR. The SFFV 5'' LTR does not interfere with in vivo expression of tk when it is flanked by homologous 5'' promoter sequences. The implication of these observations for retrovirus oncogenesis and animal cell genetics is discussed.