绿脓杆菌ATCC27853外毒素PE40基因的扩增与测序

摘要/Abstract

摘要： 目的扩增绿脓杆菌ATCC27853外毒素基因PE40，并对PCR产物进行测序鉴定，为获得用于构建分子导向药物的毒素弹头基因PE40奠定基础。方法采用PCR技术，以绿脓杆菌ATCC27853基因组DNA为模板，扩增PE40基因并测序。用DNASTAR软件将测得的序列与GenBank中的国际标准产毒株PA103的PE40基因序列进行比较。结果扩增出目的基因片断，长度为1231bp。测序表明，ATCC27853与PA103核苷酸同源性为98%，产生了7个碱基的突变，突变碱基导致第364位的天冬酰胺变成丝氨酸，第506位的丝氨酸变成色氨酸，第515位的丝氨酸变成甘氨酸。但产生变化的3个氨基酸均不是文献报道中的重要活性位点。结论产生变化的3个氨基酸不会对PEA的酶活性及细胞毒性产生很大影响，绿脓杆菌ATCC27853的PE40基因可用作分子导向药物的弹头。

关键词: 铅, 铅, 蔬菜, 根系形态

Abstract: Objective: To analyse nucleotide sequence of exotoxin A structural gene PE40 of Pseudomonas aeruginosa ATCC27853. Method: The gene PE40 was amplified from genome DNA of Pseudomonas aeruginosa ATCC2785 by PCR technique. The PCR product was purified, sequenced and compared with a standard strain PA103 from GenBank by DNASTAR software. Result: An objective gene fragment with the length 1231bp was obtained. The nucleotide sequence analysis showed that ATCC27853 shared 98% alignment with PA103. The mutations in amino acid sequence are: Asn-364 to Ser; Ser-506 to Trp and Ser-515 to Gly. Conclusion: The mutations have a little influence on the enzymatic activity and cytotoxicity, so the gene PE40 of Pseudomonas aeruginosa ATCC27853 will be usefull in the research of molecular targeted protein.

中图分类号:

Q781

顾小龙,秦建华,张宁,郭莉. 绿脓杆菌ATCC27853外毒素PE40基因的扩增与测序[J]. 中国农学通报, 2006, 22(8): 30-30.

Gu Xiaolong, Qin Jianhua, Zhang Ning, Guo Li. Amplification and Sequencing of Exotoxin A Structural gene PE40 of Pseudomonas Aeruginosa ATCC27853[J]. Chinese Agricultural Science Bulletin, 2006, 22(8): 30-30.

参考文献

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