Expression of enzymatically active reverse transcriptase in Escherichia coli.
Expression of enzymatically active reverse transcriptase in Escherichia coli.
复制标题
大肠杆菌中酶活性逆转录酶的表达。
DOI：
10.1073/pnas.82.15.4944
复制
发表时间：
1985
影响因子：
11.1
通讯作者：
Goff,SP
中科院分区：
文献类型：
--
作者：
Tanese,N;Roth,M;Goff,SP
小鼠逆转录病毒逆转录酶是一种约80,000道尔顿的单体蛋白，由病毒pol1基因的中心部分编码。为了大量制备该酶，我们构建了Moloney鼠白血病病毒trpE基因和Pol1基因部分片段的融合基因。插入的POL基因序列包括成熟酶的整个编码区和各种量的附加编码序列。这些结构中的许多都表达高水平的逆转录酶活性，即使蛋白质产物的NH2和COOH末端只接近真正蛋白质的正确末端。
Reverse transcriptase of murine retroviruses is a monomeric protein of approximately 80,000 daltons, which is encoded by the central portion of the viral pol gene. To prepare large quantities of the enzyme, we have constructed gene fusions between the trpE gene and portions of the pol gene of Moloney murine leukemia virus. The inserted pol gene sequences include the entire coding region for the mature enzyme and various amounts of additional coding sequences. Many of these constructs express high levels of reverse transcriptase activity even though the NH2 and COOH termini of the protein product only approximate the correct termini of the authentic protein.