一种新的猪囊尾蚴蛋白的cDNA分子筛选、序列分析和克隆

Screening,sequencing and cloning Sun Junmin on a cDNA encoding protein of Cysticercus cellulosae

目的免疫学筛选猪囊尾蚴cDNA文库,为猪囊尾蚴病临床免疫学诊断提供新的候选抗原分子。方法利用脑囊尾蚴病人血清免疫学筛选猪囊尾蚴cDNA文库,获得阳性克隆,经PCR技术扩增噬菌体载体中插入的cDNA片段,测序后与生物信息网站进行分析,并将一个类似血吸虫核糖体蛋白的基因克隆入PMD-18T载体。结果从cDNA文库中筛选出1个编号为5H的基因,长度是528bp;对其编码的氨基酸序列的分析显示其与日本血吸虫的L18a核糖体蛋白具有高度同源性。结论克隆了猪囊尾蚴的未知核糖体蛋白基因。

Objective To screen Taenia solium metacestodes cDNA library, and provide new candidate protein genes for clinical immunodiagnosis. Methods The cDNA library of Cysticercus cellulosae was screened by using the serum of patient with cerebral cysticercosis. After the insert of positive clone was amplified by PCP,, the nucleotide sequence of the cDNA was sequenced. The homological search of the nucleotide sequence was done by using BLASTN in NCBI. Aim gene was cloned into PMD-18T. Results A cDNA clone （named 5H） with a length of 732 bp was isolated. The clone contained one open reading flame composed of 528 bp. The predicted translation product has strong homology with ribosomal protein of Schistosoma japonicum. Conclusion A gene encoding ribosomal protein of Cysticercus ceUulosae was cloned.