论文部分内容阅读
对NCBI基因库黄龙病亚洲种病原菌序列进行拼接,并根据拼接结果在rplKAJL-rpoBC,16S/23S rRNA和omp3个位点上设计引物,采用基因组步行方法,在其亚洲种3个位点上共获得了15168bp非冗余新序列,其中,在rplKAJL-rpoBC位点上5′端上游区和3′端下游区分别获得了3218和2653bp的非冗余新序列;在16S/23S rRNA位点上5′端上游区和3′端下游区分别延伸了1853和2459bp的新序列;在omp基因位点上分别获得了4014和971bp的新序列。序列分析表明在rplKAJL-rpoBC位点上的新序列包含2个新基因(suhB和serA);16S/23S rRNA位点新序列包含4个新基因(膜转运蛋白基因、细胞壁脱氢酶假基因、5S rRNA、tRNAMet),omp位点新序列包含6个新基因(rpsB、tsf、pyrH、frr、cdsA2、cds2)。这些新序列的获得为以PCR为基础的黄龙病相关研究提供了有用的参考依据。
According to the results of splicing, primers were designed on rplKAJL-rpoBC, 16S / 23S rRNA and omp3 loci. The genomic walking method was used to amplify the sequences of three species of Asian pathogenic bacteria The 15,168bp non-redundant new sequences were obtained. Among them, 3218 and 2653bp non-redundant new sequences were obtained at the 5 ’end and 3’ end of the rplKAJL-rpoBC site respectively. On the 16S / 23S rRNA site The new sequences of 1853 and 2459bp were extended in the upstream of 5 ’end and the downstream of 3’ end respectively. New sequences of 4014 and 971 bp were obtained respectively at omp gene locus. Sequence analysis showed that the new sequence at the site of rplKAJL-rpoBC contained two new genes (suhB and serA); the new sequence of 16S / 23S rRNA site contained four new genes (membrane transporter gene, cell wall dehydrogenase pseudogene, 5S rRNA, tRNAMet). The new sequence of omp site contains 6 new genes (rpsB, tsf, pyrH, frr, cdsA2, cds2). The availability of these new sequences provides a useful reference for PCR-based studies on the yellow dragon’s disease.