摘要
通过RT-PCR扩增出捻转血矛线虫(H aem onchus contortus)氨基肽酶基因,并对该基因进行克隆、鉴定及序列测定。捻转血矛线虫氨基肽酶ORF由2 919个碱基(972个氨基酸)构成,和G enB ank中的捻转血矛线虫氨基肽酶(H 11抗原)同源性高达98%,与秀丽新杆线虫(C aenorhabd itis eleg ans)肽酶M 1家族同源性为61%,且具有氨基肽酶特征性基序HEXXH和GAM EN。将该基因亚克隆到原核表达载体并进行了表达,通过测定重组蛋白分解氨基肽酶底物的能力以及对酶抑制剂敏感性试验,进一步证实了H 11抗原具有一定的酶活性。
The cDNA gene of aminopeptidase in Haemonehus eontortus, obtained by reverse polymerase chain reaction, was cloned into pMD18-T. And then the nucleotide was sequenced and analyzed. 98% homology with H11 of Haemonchus contortus and 61% identity with peptidase M1 family of Caenorhabditis elegans were found. The HEXXH and GAMEN motifs characteristic of microsomal aminopeptidase was also got when the deduced amino acid sequence was compared with that of other Rhabditata and mammalians. To completely identification the aminopeptidase, the gene was insert into vector pET28 and expressed in E. coli DE3. Microsomal aminopeptidase activities of recombinant proteins were found and this activity could be inhibited by phenanthroline but not by phenylmethylsulfonyl floride or ethylenediaminetetraceate.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2006年第2期151-154,共4页
Chinese Journal of Veterinary Science
