Boyer P L, Ferris A L, Hughes S H
ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Maryland 21702-1201.
J Virol. 1992 Feb;66(2):1031-9. doi: 10.1128/JVI.66.2.1031-1039.1992.
We constructed a series of BspMI cassettes that simplify the introduction of specific point mutations in the polymerase domain of human immunodeficiency virus type 1 reverse transcriptase. A series of point mutants were constructed by using these cassette vectors. The RNA-dependent DNA polymerase and RNase H activities of 20 point mutations in the conserved portion of the polymerase domain were assayed. All the mutations analyzed are conservative substitutions of evolutionarily conserved amino acids. The mutations were divided into four classes. The first class has little effect on either polymerase or RNase H activity. The second class affects RNase H but not polymerase activity, while the third class has a normal RNase H activity with diminished polymerase activity. The fourth class affects both activities.
我们构建了一系列BspMI盒式载体，这些载体简化了在人类免疫缺陷病毒1型逆转录酶聚合酶结构域中引入特定点突变的过程。利用这些盒式载体构建了一系列点突变体。对聚合酶结构域保守部分的20个点突变的RNA依赖性DNA聚合酶和核糖核酸酶H活性进行了测定。所有分析的突变都是进化上保守氨基酸的保守替换。这些突变分为四类。第一类对聚合酶或核糖核酸酶H活性几乎没有影响。第二类影响核糖核酸酶H但不影响聚合酶活性，而第三类具有正常的核糖核酸酶H活性但聚合酶活性降低。第四类影响这两种活性。
