Mutagenesis and Cloning of HIV-1 Protease Gene by Polymerase Chain Reaction

RUN Xiao-Jun; SU Cheng-Zhi; JI Chang-Hua

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April 1993

RUN Xiao-Jun, SU Cheng-Zhi and JI Chang-Hua. Mutagenesis and Cloning of HIV-1 Protease Gene by Polymerase Chain Reaction[J]. Virologica Sinica, 1993, 8(2).

Citation: RUN Xiao-Jun, SU Cheng-Zhi, JI Chang-Hua. Mutagenesis and Cloning of HIV-1 Protease Gene by Polymerase Chain Reaction .VIROLOGICA SINICA, 1993, 8(2) : 154.

用PCR突变技术克隆艾滋病病毒蛋白酶基因

1.
第四军医大学生化教研室
2.
第四军医大学生化教研室西安

摘要

作者设计并合成了一对用于PCR技术的突变引物HIV-1 Pr1和HIV-1Pr2,分别在两引物中设计了两个突变点,使突变后基因含有EcoRI、HindⅢ和TAA序列,便于HIV-1 Pr基因的定向克隆和表达。用HIV-1 Pr1和HIV-1 Pr2作引物,采用PCR方法从HIV-1基因组DNA中扩增出了一个360bp长的DNA片段,用EcoRI和HindⅢ双酶切法将此片段定向克隆入pUC19质粒,将克隆基因插入M13mp18进行DNA序列分析。结果表明,该基因序列的读框完全正确,从而为HIV-1 Pr基因的表达及抑制剂的研究奠定了基础。
- 人免疫缺陷病毒
- , 聚合酶链反应
- , 基因克隆
- , 基因突变

Mutagenesis and Cloning of HIV-1 Protease Gene by Polymerase Chain Reaction

1.
Department of Biochemistry
2.
The Fourth Military Medical University

Abstract

In the present work, the primers HIV-1 Pr1 and HIV-1 Pr2 were synthesized in which the EcoRI and Hind Ⅲ recognition sites as well as the termination codons were designed for convenience of subsequent cloning and expression Micrograms of target gene sequence was produced by 30 rounds of amplification with pJG423 as a template The amplified gene was digested with both EcoRI and Hind Ⅲ and ligated to pUC19 predigested with the same restriction endonucleases. The recombinant plasmid pUPR1 nd pUPR2 were identifi...
- Human immunodeficiency virus Gene cloning Polymerase chain reaction Gene mutagenesis

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Mutagenesis and Cloning of HIV-1 Protease Gene by Polymerase Chain Reaction

1. Department of Biochemistry
2. The Fourth Military Medical University

Abstract: In the present work, the primers HIV-1 Pr1 and HIV-1 Pr2 were synthesized in which the EcoRI and Hind Ⅲ recognition sites as well as the termination codons were designed for convenience of subsequent cloning and expression Micrograms of target gene sequence was produced by 30 rounds of amplification with pJG423 as a template The amplified gene was digested with both EcoRI and Hind Ⅲ and ligated to pUC19 predigested with the same restriction endonucleases. The recombinant plasmid pUPR1 nd pUPR2 were identifi...