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昆虫病原线虫共生菌YL001菌毛蛋白亚基基因的克隆与序列分析
冯纪年1, 李玉凤1, 王永宏2
1.西北农林科技大学 植保资源与病虫害防治教育部重点实验室;2.西北农林科技大学 无公害农药研究服务中心
摘要:
为了准确克隆昆虫病原线虫共生菌YL001菌毛蛋白亚基基因,根据已报道的菌毛蛋白亚基序列设计引物,以昆虫病原线虫共生菌YL001的总DNA为模板进行PCR扩增,获得1条大约500 bp的特异性片断,将此片断克隆到T-easy载体上,获得了重组子pGEM-PSYL001,序列测定和结构分析结果表明,PSYL001阅读框全长537bp,编码179个氨基酸,预测分子量和等电点分别为18.9 ku和6.94,GenBank登录号为DQ537944。与GenBank(AY140909)上公布的Xenorhabdus nematophilus菌毛蛋白亚基核苷酸序列的同源性为99%,氨基酸序列的同源性为98%。上述分析表明,菌毛蛋白亚基基因得到了成功克隆。
关键词:  昆虫病原线虫共生菌  菌毛蛋白亚基  基因克隆  序列分析
DOI:
分类号:
基金项目:国家“十五”科技攻关项目(2002BA516A04);西北农林科技大学研究生教育创新计划项目(05YCH007)
Cloning and characterization analysis of Pilin Subunit gene from Xenorhabdus nematophilus YL001
Abstract:
In order to clone the gene of Pilin Subunit from Xenorhabdus nematophilus YL001 accurately,a pair of primers were designed based on the sequence of Pilin Subunit reported previously.A Specific band (about 500 bp in length)was amplified from DNA of Xenorhabdus nematophilus YL001.The segment was cloned into T-easy vector.Results of sequencing and structural analysis show that the length of PSYL001 is 537 bp,and 179 amino acids are encoded.The predicted MW and pI are 18.9 ku and 6.94.The accession number in GenBank is DQ537944.Comparison results show that this gene has 99% similarity to nucleotides level and 98% similarity to amino level according to AY140909,which is Pilin Subunit gene from Xenorhabdus nematophilus in GenBank.The analysis indicates that the Pilin Subunit gene has been cloned successfully.
Key words:  Xenorhabdus nematophilus YL001  Pilin Subunit  gene cloning  characterization analysis