短小芽胞杆菌抑菌相关基因的高效克隆  

Highly efficient cloning of functional genes relating to antimicrobial activity of Bacillus pumilus

作　　者：刘通[1] 陈云鹏[1] 李琼洁 顾振芳[1] 

机构地区：[1]上海交通大学农业与生物学院,农业部都市农业(南方)重点实验室,上海200240

出　　处：《植物保护学报》2014年第4期423-428,共6页Journal of Plant Protection

基　　金：国家自然科学基金(31071721)

摘　　要：为有效挖掘生防短小芽胞杆菌Bacillus pumilus的抑菌相关基因,对DX01菌株进行了Solexa基因组扫描,并选取其中2个代表性基因TasA和Surf2,以DX01基因组DNA为模板,采用PCR方法对其进行克隆、序列比对及系统进化分析。结果表明,DX01菌株共得到4267个基因,有3145个具有明确生物学功能的蛋白。其中,与短小芽胞杆菌抑菌活性相关的基因(簇)有84个,包括2个细菌防御酶基因、2个细菌生物膜形成相关基因、1个信号转导基因、27个细胞壁降解或水解酶基因、15个抗生素合成或转运基因、4个抑菌蛋白基因、1个次生代谢物合成基因簇及32个细菌鞭毛生物合成与调控相关基因。TasA基因在芽胞杆菌属细菌中具有高度的保守性,各同源基因的序列相似性在92%以上;而Surf2基因的同源基因在该属细菌中具有一定的变异性,有的序列相似性低至73%。The genome sequencing of Bacillus pumilus DX01 was performed with Illumina /Solexa Hiseq2000 to identify the bacterial functional genes encoding antimicrobial activities. A total of 4267 genes and3 145 protein-coding sequences with clear biological functions were predicted using bioinformatic software combined with retrieval of the public protein database of NCBI. Eighty-four functional genes relating to antimicrobial activities were found in the genome of B. pumilus. Among them,there were two bacterial defense enzyme genes,two genes relating to bacterial biofilm formation,one signal transduction gene,twenty-seven cell wall degradation and hydrolase genes,fifteen genes relating to antibiotic synthesis or transport,four antimicrobial protein genes,one secondary metabolite synthesis gene cluster,and thirtytwo bacterial flagellar biosynthesis and regulation genes. Finally,in order to confirm the precision of the assembled sequences of B. pumilus strain DX01 Solexa sequencing,two representative antimicrobial genes,TasA and Surf2,were selected for cloning with PCR technique. The TasA gene was found to share over 92% similarity in DNA sequence with its homologous genes from other Bacillus strains and be highly conserved in gene evolution based on DNA sequence alignment and phylogenetic analyses. However,homologous genes of Surf2 had some variation in Bacillus and some DNA sequence similarities were as low as 73% in alignment analysis.

关 键 词：短小芽胞杆菌 Solexa扫描 抑菌相关基因 

分 类 号：S476[农业科学—农业昆虫与害虫防治]