Reverse transcriptase - The use of cloned moloney murine leukemia virus reverse transcriptase to synthesize DNA from RNA
Reverse transcriptase - The use of cloned moloney murine leukemia virus reverse transcriptase to synthesize DNA from RNA
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DOI：
10.1007/bf02762340
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发表时间：
1997-08-01
影响因子：
2.6
通讯作者：
DAlessio, JM
中科院分区：
文献类型：
--
作者：
Gerard, GF;Fox, DK;DAlessio, JM
逆转录酶（RT）是将RNA转化为DNA所需的关键酶。莫洛尼鼠白血病病毒（MMLV）RT的克隆使得能够工程化缺乏内源性RNA酶H活性的RT。RT在没有RNase H的情况下更有效地催化cDNA合成。我们在这里描述的MMLV RT和RNase H-减MMLV RT没有在一个单一的位置在其他地方总结的一些属性，提供了一个基础，最好地利用这些酶在cDNA合成。此外，还提供了在单管双链cDNA合成、[P-32]cDNA合成以及RT-PCR和长RT-PCR中使用MMLV RT的一般指南和详细方案。
Reverse transcriptase (RT) is the key enzyme required for conversion of RNA to DNA. Cloning of Moloney murine leukemia virus (MMLV) RT has enable engineering an RT that lacks endogenous RNase H activity. RT catalyzes cDNA synthesis more efficiently in the absence of RNase H. We describe here a number of properties of MMLV RT and RNase H-minus MMLV RT not summarized in a single location elsewhere, providing a basis for best use of these enzymes in cDNA synthesis. In addition, general guidelines and detailed protocols are provided for use of MMLV RTs in one tube double-stranded cDNA synthesis, in [P-32]cDNA synthesis, and in RT-PCR and long RT-PCR.