[嗜肺军团菌PAL基因的克隆与表达]

[Cloning and expression of PAL gene of Legionella pneumophila].

作者信息

Tian Yu, Chen Jian-ping, Yang Chun-lei, Liu Ming-jie

机构信息

Department of Parasitology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2007 Jul;38(4):583-6, 598.

PMID:17718416

Abstract

OBJECTIVE

To develop DNA vaccine for Legionella pneumophila.

METHODS

PAL gene of Legionella pneumophila was amplified with PCR. The amplified DNA was ligated to pcDNA3.1(+) vector. The recombinant plasmid, which was identified by restriction analysis, PCR and sequence analysis, was named pcDNA3.1-PAL. The NIH3T3 cells were transfected with the recombinant plasmid pcDNA3.1-PAL by Lipofection. Transient and stable expression products of the PAL gene were detected by immunofluorescence and RT-PCR.

RESULTS

The recombinant plasmid pcDNA3.1-PAL expressed PAL protein in the eukaryotic cell NIH3T3.

CONCLUSION

This study has built a foundation for the development of PAL gene DNA vaccine for Legionella pneumophila.

摘要

目的

研发嗜肺军团菌DNA疫苗。

方法

采用聚合酶链反应（PCR）扩增嗜肺军团菌的PAL基因。将扩增的DNA连接到pcDNA3.1(+)载体上。经限制性酶切分析、PCR及序列分析鉴定后的重组质粒命名为pcDNA3.1-PAL。采用脂质体转染法将重组质粒pcDNA3.1-PAL转染至NIH3T3细胞。通过免疫荧光和逆转录-聚合酶链反应（RT-PCR）检测PAL基因的瞬时和稳定表达产物。