羊源类鼻疽伯克霍尔德菌BPSS1512基因克

发布时间：2018-06-02 04:38

  本文选题：类鼻疽伯克霍尔德菌 + BPSS基因　； 参考：《中国畜牧兽医》2017年08期

【摘要】：为克隆羊源类鼻疽伯克霍尔德菌BPSS1512基因,并对其编码的蛋白进行生物信息学分析,以类鼻疽伯克霍尔德菌基因组为模板,参照GenBank中Burkholderia pseudomallei K96243株基因组DNA序列(登录号:NC_006351.1)设计引物,PCR扩增BPSS1512基因,构建重组质粒,SDS-PAGE和Western blotting分析其蛋白表达,DNAMAN等软件对BPSS1512基因编码的氨基酸序列进行分析。结果显示,PCR扩增成功得到1 425bp的特异性条带,BamHⅠ和HindⅢ双酶切后得到约为5 000和1 500bp的条带,表明重组质粒pET-28a-BPSS1512构建成功,IPTG浓度为10mmol/L,诱导时间8h为最适宜的诱导条件。BPSS1512基因编码的蛋白质分子质量为53ku,在包涵体中表达;在BPSS1512蛋白二级结构中,α-螺旋、延伸链和无规卷曲分别占24.05%、14.77%、61.18%,并且疏水性区域分布在-2.0~+2.4之间,说明BPSS1512蛋白具有较强的疏水性,本试验结果可为类鼻疽病的防制提供参考依据。
[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，

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