鸡法氏囊B淋巴细胞中与鸡传染性法氏囊病病毒VP2相互作用蛋白筛选

Screening proteins interacting with infectious bursa disease virus Gt VP2 from cDNA library of B lymphoid cells of the bursa of fabricius

摘要:

为了获得鸡法氏囊B淋巴细胞中与鸡传染性法氏囊病病毒 (IBDV) VP2相互作用的蛋白质，利用酵母双杂交系统，用IBDV VP2蛋白为诱饵蛋白，筛选鸡法氏囊B淋巴细胞cDNA表达文库。将表达文库质粒转化含IBDV VP2诱饵质粒的酵母感受态细胞，检测报告基因在相应的营养缺陷型培养基 (SD/-Leu/-Trp/-His) 上表达情况，进一步经β-半乳糖苷酶报告基因检测，筛选到16个阳性克隆。提取阳性克隆质粒，经测序分析获得5个原鸡基因序列，分别是：线粒体DNA、蛋白质O位N-乙酰葡萄糖胺糖基化转移酶、肿瘤

Abstract:

To screen the interactive proteins with IBDV Gt VP2 protein from cDNA library of B Lymphoid cells of the bursa of fabricius. The expression cDNA library plasmids was transformed to the yeast competent cells, which have the bait plasmid-Gt VP2. After testing for growth in synthetic complete medium lacking histidine and uracil and for production of b-galactosidase (X-gal), we obtained 16 positive clones. We searched the gene sequences of positive clones in the NCBI website. The blast results showed that five positive clones were the gallus sequences. They were Gallus gallus breed mitochondrial DNA, O_GlcNAc transferase, Tumor protein p53 binding protein, Stathmin and Chondroitin sulfate GalNAcT-2, respectively. This study is helpful for the further identifying the receptors of IBDV in B Lymphoid cells of the bursa of fabricius.