日本血吸虫酪蛋白激酶Ⅱβ亚基的序列分析

Cloning and sequence analysis on casein kinase Ⅱ beta subunit of sohistosoma japonicum

目的 发现并克隆日本血吸虫新基因。方法 对已获得的日本血吸虫表达序列标签 (EST)进行同源性分析 ,识别血吸虫新基因 ;根据EST设计引物 ,用 3′RACE从mRNA中扩增获得新基因全长 ,用生物信息学技术对获得的编码基因进行结构与功能分析 ,将新基因的完整编码阅读框克隆入原核表达载体 pGEX -4T -1。 结果 用 3′RACE获得一个EST的 3′端所缺序列 ,得到完整编码阅读框 ,其开放阅读框 (ORF) 654bp ,编码 2 17个氨基酸。利用生物信息学技术确定其为日本血吸虫酪蛋白激酶Ⅱβ亚单位的编码基因。重组质粒经双酶切及测序鉴定 ,证明日本血吸虫CKⅡβ原核表达质粒构建成功。 结论 扩增并成功克隆日本血吸虫酪蛋白激酶Ⅱβ亚单位编码基因的全长cDNA 。

Objective To recognize and clone the novel genes of Schis tosoma japonicum(Sj).2糤T5”HZ]MethodsMethods The ESTs obtained in our laboratory were analyzed by homologous searching,the novel genes were recognized.According to the intere sted EST,the full-length cDNA of the novel gene was obtained by 3′ RACE PCR .Th e cDNA sequence were cloned into pGEX-4T-1 vector and analyzed by bioinformatics . Results A new cDNA sequence was obtained,the sequence con tains a full-length sequence with 654bp ORF,which encoded 217 amino acid residue s.The new gene was identified to be the gene encoding casein kinase Ⅱ beta subu nit.The sequences were cloned into pGEX-4T-1 vector and identified by restrictio n analysis and sequencing.Conclusion The full-length cDNA sequences encoding Sj cas ein kinase Ⅱ beta subunit were firstly sequenced and cloned,which give the basi s for further study.