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苏云金芽胞杆菌以色列亚种20kDa蛋白质对CytA蛋白溶细胞作用的影响

The Influence of the 20kDa Protein from Bacillus thuringiensis subsp. israelensis on the Cytolytic Activity of CytA

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【作者】 刘子铎孙明陈亚华喻子牛R.ManassherobE.Ben-DovA.Zaritsky

【Author】 LIU Zi-Duo; SUN Ming; CHEN Ya-Hua; YU Zi-Niu; R. Manassherob; E. Ben-Dov; A.Zaritsky ( Dept. of Microbial Science and Technology Huazhong Agricultural University Wuhan 430070) ( Dept. of Life Sciences Ben-Gurion University of the Negev Israel)

【机构】 华中农业大学农业微生物农业部重点实验室微生物科学技术系!武汉430070华中农业大学农业微生物农业部重点实验室微生物科学技术系!武

【摘要】 为了证明苏云金芽胞杆菌以色列亚种20kDe蛋白质对CytA蛋白溶细胞作用的影响, 根据20kDe蛋白质和cytA蛋白基因的核苷酸序列,用AMPLIFY程序设计了一套带有酶切位 点的引物,经PCR扩增分别获得了20kDe蛋白质和cytA蛋白基因。将其基因与表达载体 pUHE24连接并转化到大肠杆菌XLI和DHS 分别获得含20kDa蛋白质基因的克隆子 LZ29;含cytA基因的克隆子LZcytA和含有二者基因的重组子LZ20A.在IPTG诱导下,测定 了不同克隆株基因表达产物对大肠杆菌细胞生长的影响。结果表明:LZ20的细胞生长不受影 响;LZcytA的细胞被杀死;LZ20A的细胞生长也不受影响。这表明20kDa蛋白质基因与cytA 蛋白基因重组后,20kDa蛋白质基因表达产物可保护CytA蛋白对大肠杆菌的溶细胞作用,而 巳这种作用并不因不同大肠杆菌受体而改变。

【Abstract】 In order to understand the influence of the 20kDa protein on the cytolytic activity of CytA protein, a set of PCR primers were designed to amplify 20kDa protein and CytA protein genes. The genes were liga0ted to E. colt expression vector pUHE24 and transformed into E coli XLI and DH5. The clones, LZcytA, and LZ20A containing 20kDa protein gene, cytA protein gene and 20kDa-cytA genes were obtained respectivelsy. The influence of the clones on the growth of E. coli scells was determined under induction of IPTG. The results showed that the growth of LZ20 cellss were not affected, LZcytA cells were killed, and the growth of LZ20A cells were not affected. It was suggested that expression product of 20kDa protein gene protected the host cells from the cytolytic effect of CytA protein. This was supported by using different host strains.

  • 【文献出处】 遗传学报 ,ACTA GENETICA SINICA , 编辑部邮箱 ,1999年01期
  • 【分类号】Q347
  • 【被引频次】14
  • 【下载频次】44
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