黑胸大蠊若虫免疫兔血清对美洲大蠊若虫cDNA表达文库的筛选及克隆分析

目的从美洲大蠊若虫cDNA文库中筛选黑胸大蠊抗原相关基因,以期获得黑胸大蠊有潜在药用价值的蛋白质、多肽成分。方法用黑胸大蠊若虫免疫兔血清筛选美洲大蠊若虫cDNA文库,对阳性克隆进行PCR鉴定和序列测定,通过国际互联网进行核苷酸序列的同源性分析及编码蛋白质的结构分析和功能预测。结果经三轮复筛,从多个持续阳性克隆中挑选出3个进行测序、分析,其中N1克隆为新基因,Score<50,命名为Parcxpwnx01,GenBank登录号为AY838802。该基因编码231个氨基酸,初步预测该蛋白可能为一具信号转导功能的跨膜蛋白。N2克隆与美洲大蠊核蛋白体16SRNA基因高度同源,N3与黑胸大蠊线粒体DNA高度同源。结论免疫筛选获得与黑胸大蠊若虫抗原相关的基因克隆,其编码的蛋白结构和功能有待进一步研究。

Immunoscreening of the Periplaneta americana Nymph cDNA Library with Periplaneta fulignosa Immunized Rabbit Sera

Objective To clone the genes of the antigens of Periplaneta americana. Method Periplaneta Americana nymph cDNA library was screened with Periplaneta fulignosa immunized rabbit sera. Positive clones were identified by PCR, nucleotide sequencing and data analysis through BLASTn software and Expert Protein Analysis System of Swiss Institute of Bioinformatics. Result After the third screening, three positive clones were analyzed. N1 is a novel gene (GenBank accession number: AY838802) and encodes a protein consisting of 231 amino acids, which is predicted to be a transmembrane protein with a function of signal transduction. N2 and N3 are highly homologous to Periplaneta americana 16S ribosomal RNA gene and Periplaneta fuliginosa mitochondrial DNA, respectively. Conclusion Their protein structures and functions remain to be studied.