牛干扰素—γ基因的克隆、序列分析及其表达产物的生物学活性的研究.pdf

摘 要 摘 要 铷 根据GenBank上发表的牛干扰素丫基因序列设计引物。从牛外周血淋巴细胞中提取 -气 电泳显示牛干扰素丫扩增片段约为500bp。序列分析结果表明，牛干扰素1，基因序列全 在约19．4kd处有表达蛋白带，表达产物经免疫荧光染色鉴定为阳性。生物学活性研究 表明：重组牛Y一干扰素对牛外周血淋巴细胞具有抑制其分裂及抗其凋亡的作用。 h 关键词：牛，干扰素一Y，R卜PCR，序列分析，克隆，表达，细胞分裂，细胞调亡 ～ Abstract Abstract Basedonthe nucleotide ofbovine of published sequence pair RT-PCRwere and from bloodT primersdesigned RNA，isolated synthesized．Totle pefipheral cellsof usedas to DNAreverse bovine，was templategeneratecomplementaryby DNA were chain transcription．The reaction． 500bp fragmentamplified bypolymerase resultsdemonstratedthat contains51 a501 Sequencing interferon--),gene7bp、析th bpopen a of166 aminoacidewhosemolecularis1 readingfragment(OR．F)encoding peptide 9．4kd， whichis accordwithreference into completely array．Thecloning Wasinserted genefragment vector when werecut BamHI／EcoRI．Therecombinant expressionpBV220they by plasmid wastransformedintoDH5astrain SDS．PAGEresultshowed byCaCl2．The thatcloned recombinantw