本文選題：類鼻疽伯克霍爾德菌 + BPSS基因��； 參考：《中國畜牧獸醫(yī)》2017年08期

【摘要】：為克隆羊源類鼻疽伯克霍爾德菌BPSS1512基因,并對其編碼的蛋白進(jìn)行生物信息學(xué)分析,以類鼻疽伯克霍爾德菌基因組為模板,參照GenBank中Burkholderia pseudomallei K96243株基因組DNA序列(登錄號:NC_006351.1)設(shè)計(jì)引物,PCR擴(kuò)增BPSS1512基因,構(gòu)建重組質(zhì)粒,SDS-PAGE和Western blotting分析其蛋白表達(dá),DNAMAN等軟件對BPSS1512基因編碼的氨基酸序列進(jìn)行分析。結(jié)果顯示,PCR擴(kuò)增成功得到1 425bp的特異性條帶,BamHⅠ和HindⅢ雙酶切后得到約為5 000和1 500bp的條帶,表明重組質(zhì)粒pET-28a-BPSS1512構(gòu)建成功,IPTG濃度為10mmol/L,誘導(dǎo)時間8h為最適宜的誘導(dǎo)條件。BPSS1512基因編碼的蛋白質(zhì)分子質(zhì)量為53ku,在包涵體中表達(dá);在BPSS1512蛋白二級結(jié)構(gòu)中,α-螺旋、延伸鏈和無規(guī)卷曲分別占24.05%、14.77%、61.18%,并且疏水性區(qū)域分布在-2.0~+2.4之間,說明BPSS1512蛋白具有較強(qiáng)的疏水性,本試驗(yàn)結(jié)果可為類鼻疽病的防制提供參考依據(jù)。
[Abstract]:The BPSS1512 gene of Borkhold-like bacteria from cloned sheep was analyzed by bioinformatics. The genome of Burkhold-like bacillus mallei was used as a template for bioinformatics analysis. According to the genomic DNA sequence of Burkholderia pseudomallei K96243 strain in GenBank (accession number: NCSTX 006351.1), the BPSS1512 gene was amplified by PCR. The recombinant plasmid was constructed by SDS-PAGE and its protein expression was analyzed by Western blotting. The amino acid sequence of BPSS1512 gene was analyzed. The results showed that the specific bands of 1 425bp, BamH 鈪，

本文編號：1967438