D型产气荚膜梭菌贵州分离株ε毒素的DNA序列测定与分析

Detection and Analysis of Nucleotide Sequence of Epsilon-toxin Gene of Clostridium perfringens Type D

为研究产气荚膜梭菌ε毒素的结构与功能,以及对该病的防治提供基础依据,利用D型产气荚膜梭菌贵州分离株（CP02株）,根据GenBank中登记的产气荚膜梭菌ε毒素基因设计特异性引物进行PCR扩增,扩增产物纯化后测定核苷酸序列并与NCBI登录的参考毒株进行相似性比较。结果表明：D型产气荚膜梭菌CP02株基因片段大小为456bp,与CWD_CN_409、NCTC_8533、NCTN_6121、ETX21512、KurCpD1、CtrCpD2、IVRI_Vac1和IVRI_49菌株的核苷酸序列相似性在98.5%~100%,推导的氨基酸序列相似性在98.4%~100%。碱基突变以A-G、C-T间的转换为主,也发生低频率的A-C、G-T间颠换,但突变均为点突变。

To study the structure and function of epsilon-toxin gene of C.perfringens,and provide a fundamental basis for controlling the disease, the special primers of epsilon-toxin gene within C. perfringens were designed for PCR amplification according to the accession in GenBank by using the isolated strain CP02,nucleotide sequence was detected after purification of the PCR products and was compared with the NCBI reference sequences.Results：The gene segment size of CP02 was 456 bp,the rate of nucleotide sequence similarity between the tested strain and strain CWD_CN_409,NCTC_8533,NCTN_6121,ETX21512,KurCpD1,CtrCpD2,IVRI_Vac1 and IVRI_49 were 98.5%~100% and similarity of the deduced amino acid sequence were 98.4% ~ 100%.The base mutation was mainly the transition between A-G and C-T,which occurred between A-C and G-T in low frequency,but the mutation were all point mutation.