日本血吸虫中国大陆株基因重组抗原Sj 22.6kDa的核苷酸序列分析  

RESEARCHES ON THE NUCLEOTIDE SEQUENCING OF THE RECOMBINANT Sj 22 6 kDa ANTIGEN GENE OF SCHISTOSOMA JAPONICUM *

作　　者：张桂筠[1] 张兆松[1] 陈淑贞[1] 沈一平[1] 吴海玮[1] 苏川[1] 王荣芝[1] 吴观陵[1] 

机构地区：[1]南京医科大学寄生虫学教研室

出　　处：《中国寄生虫学与寄生虫病杂志》1998年第2期105-108,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国务院总理专项研究基金

摘　　要：目的：对ｐＧＳｊ２４克隆化基因进行核苷酸序列分析，了解其编码蛋白的属性。方法：常规制备ｐＧＳｊ２４克隆化基因并重组入测序载体Ｍ１３ｍｐ１９，以ＤＹＥＰＲＩＭＥＲ荧光测序试剂盒进行核苷酸序列测定。分别以ＤＮＡＳＩＳ和ＧＯＬＤＫＥＹ软件对序列资料进行分析。结果：ｐＧＳｊ２４克隆化基因长８４０ｂｐ，含一开放阅读框，可编码一分子量为２２．６ｋＤａ的蛋白质。开读框上游和下游均有终止密码子。该基因与已发表的日本血吸虫２２．６ｋＤａ蛋白的编码基因同源性达９５％，编码区同源性达９９．７％。在该基因内有一段典型的ＥＦ－Ｈａｎｄ钙结合区序列，并有内质网导肽、微体导向信号等功能位点。预测该蛋白质内可能的抗原决定簇位置为第２９－３２、６３－６８和８７－１０１等氨基酸片段。结论：ｐＧＳｊ２４克隆化基因为日本血吸虫２２．６ｋＤａ抗原编码基因。AIM: To sequence the cloned gene in pG Sj 24 and to identify the encoded protein. METHODS: The cloned gene in pG Sj 24 was digested from the recombinant plasmid by EcoRI,ligated into the M13mp19 vector and sequenced by automatic sequencer.The sequence was analyzed by Goldkey DNA and Protein Analytical Program and DNASIS Program.RESULTS: The pG Sj 24 cloned gene was demonstrated to be 840 bp containing one opened reading frame (ORF) with an initiation codon ATG at position 23 nt and a termination codon TAA at position 596 nt, encoding a protein with a molecular weight of 22 6 kDa.At the upstream and downstraem of the ORF there were termination codons,so the encoded protein was unable to be larger.However, there was a termination codon TAA at position 11 nt,suggesting why the 22 6 kDa protein expressed separately.The nucleotide sequence of the pG Sj 24 cloned gene shared 95% identity with that of the corresponding part of S.japonicum 22 6 kDa antigen gene,and 99 7% identity in the encoding part.The deduced amino acid scquence analysis showed a sequence motif known as EF Hand calcium binding domain, several endoplasmic reticulum targetting sequences and microbodies C terminal targeting signals.The possible antigen determinants were predicted within the amino acid fragments of 29-32aa,63-68aa and 87-101aa. CONCLUSIONS: The cloned gene in pG Sj 24 is the gene that encodes Sj 22 6 kDa antigen.

关 键 词：日本血吸虫 重组抗原 中国大陆株 核苷酸 序列 

分 类 号：R383.24[医药卫生—医学寄生虫学]