流产型布鲁氏菌Omp22基因酵母双杂交诱饵载体的构建及鉴定

›› 2011, Vol. 38 ›› Issue (6): 83-86.

流产型布鲁氏菌Omp22基因酵母双杂交诱饵载体的构建及鉴定

李向阳^1,3, 张嘉保²

1. 吉林大学农学部畜牧兽医学院, 吉林长春 130062;2. 吉林大学农学部实验动物中心, 吉林长春 130062;3. 内蒙古民族大学成人教育学院, 内蒙古通辽 028043

收稿日期:2010-11-25 修回日期:1900-01-01 出版日期:2011-06-20 发布日期:2011-06-20

Construction and Identification of Bait Vector Containing Omp22 Gene from Brucella abortus in Yeast Two Hybrid System

LI Xiang-yang^1,3, ZHANG Jia-bao²

1. College of Animal Science and Veterinary Medicine, Jilin University,Changchun 130062, China;2. Laboratory Animal Center, Jilin University, Changchun 130062, China;3. Inner Mongolia University for the Nationalities, Tongliao 028043, China

Received:2010-11-25 Revised:1900-01-01 Online:2011-06-20 Published:2011-06-20

摘要/Abstract

摘要： 通过PCR获得流产型布鲁氏菌Omp22基因的编码区,将其克隆到pSOS载体中,构建酵母双杂交系统的诱饵载体pSOS-Omp22。测序正确后,将重组质粒导入cdc25H检测其表达产物对酵母细胞有无毒性和自激活作用,且在酵母细胞中定位是否正确。结果表明,获得了正确的流产型布鲁氏菌Omp22基因编码区,并成功克隆到pSOS诱饵载体中,且转化有诱饵载体的cdc25H在SD/Glucose(-L)营养缺陷平板上生长良好,说明表达产物对酵母细胞无毒性,对报告基因也无自激活作用且其定位正确。表明pSOS-Omp22可应用在酵母双杂交系统中,用于寻找巨噬细胞cDNA文库中与Omp22相互作用的蛋白质。

关键词: 酵母双杂交系统; Omp22基因; 诱饵载体; 自激活

Abstract: The coding region of Omp22 gene from Brucella abortus was amplified by PCR and then fused with pSOS vector. After confirmation with sequence analysis, the plasmid was transformed into the yeast cell cdc25H, the toxicity and transcriptional auto activation of expressed protein were tested. The results indicated that the coding region was successfully amplified and subcloned into pSOS vector, and the cdc25H transformed with bait plasmid grew well on SD/glucose (-L) plate. These data showed that the bait vectors pSOS-Omp22 was expressed correctly without toxicity and could not auto activate. The pSOS-Omp22 could be applied for the screening of proteins interacting with the Omp22 in macrophages cDNA library in the yeast two hybrid systems.

Key words: yeast two hybrid system; Omp22 gene; bait vector; self-activation

中图分类号:

S852.61⁺4

李向阳;张嘉保. 流产型布鲁氏菌Omp22基因酵母双杂交诱饵载体的构建及鉴定[J]. , 2011, 38(6): 83-86.

LI Xiang-yang;ZHANG Jia-bao. Construction and Identification of Bait Vector Containing Omp22 Gene from Brucella abortus in Yeast Two Hybrid System[J]. China Animal Husbandry & Veterinary Medicine, 2011, 38(6): 83-86.

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