根据多房棘球绦虫emY162基因序列设计引物,利用PCR以细粒棘球绦虫原头蚴和成虫的基因组DNA和cDNA为模板扩增egG1Y162基因,构建PUCm-T/egG1Y162和PUCm-T/egG1Y162 cDNA重组质粒,经PCR、酶切及测序后,进行序列分析。细粒棘球绦虫2个不同发育阶段均克隆出egG1Y162基因,从基因组DNA和cDNA克隆得到的片段长度分别为1 680 bp和459 bp,登录号分别为AB458258和AB458259。
A pair of primers (egG1Y162) were designed according to the nucleotide sequence of Echinococcus multilocularis emY162 antigen gene. Using genomic DNA and cDNA from protoscoleces and adult worms of E. granulosus as templates, PCR was performed with the primers to obtain fragments of egG1Y162 gene. PUCm-T/egG1Y162 recombinant plasmids and PUCm-T/egY162 cDNA recombinant plasmids were constructed and identified by PCR, digestion with restriction enzyme and sequencing. The egG1Y162 antigen gene was amplified in protoscoleces and adult worms of E. granulosus. The size of the egG1Y162 gene was 1 648 bp and cDNA was 459 bp, and GenBank accession numbers were AB458258 and AB458259, respectively.