CAJ | 학술논문

目的研究中国炭疽芽胞杆菌(炭疽杆菌)荚膜质粒基因单核苷酸多态性(SNP)特征.方法选择中国不同分离年代、地点和来源的95株炭疽杆菌,采用PCR方法扩增荚膜质粒基因上的23个SNP位点,然后进行测序并进行聚类分析.结果通过聚类分析,95株炭疽杆菌可分为5个群,23个SNP位点中17个位点相同,6个位点存在多态性,其中17.89％(17/95)的菌株与参考菌株Pastuer同源,38.95％(37/95)的菌株与参考菌株Ames Ancestor同源,其余菌株则不同于目前已知的全基因组测序菌株；3株菌在PS-34位点扩增基因片段中缺失长度约80bp的序列,待测的SNP位点包括在该缺失片段中；9株菌株在所有SNP位点扩增阴性,经炭疽杆菌荚膜质粒基因特异引物扩增证实这些菌株缺失荚膜质粒基因.结论中国炭疽杆菌荚膜质粒SNP位点具有遗传稳定性和自身的特异性,6个SNP位点可作为基因分型的指标.
목적 연구중국탄저아포간균(탄저간균)협막질립기인단핵감산다태성(SNP)특정.방법 선택중국불동분리년대、지점화래원적95주탄저간균,채용PCR방법확증협막질립기인상적23개SNP위점,연후진행측서병진행취류분석.결과 통과취류분석,95주탄저간균가분위5개군,23개SNP위점중17개위점상동,6개위점존재다태성,기중17.89％(17/95)적균주여삼고균주Pastuer동원,38.95％(37/95)적균주여삼고균주Ames Ancestor동원,기여균주칙불동우목전이지적전기인조측서균주；3주균재PS-34위점확증기인편단중결실장도약80bp적서렬,대측적SNP위점포괄재해결실편단중；9주균주재소유SNP위점확증음성,경탄저간균협막질립기인특이인물확증증실저사균주결실협막질립기인.결론 중국탄저간균협막질립SNP위점구유유전은정성화자신적특이성,6개SNP위점가작위기인분형적지표.
Objective To study the characteristic of single nucleotide polymorphism (SNP)in capsule plasmid gene of Bacillus anthracis isolated from China.Methods 95 Bacillus anthracis isolates from different sources were selected.23 SNP sites were amplified by PCR method,sequenced and analyzed by clustering analysis.Results 95 Bacillus anthracis isolates were divided into 5 groups by cluster analysis.The identified isolates had the same sequence features in 17 sites and different nucleotide sequence in the other 6 sites of the 23 SNP sites.17.89％ (17/95) of the isolates had homologous locus sequences compared with the reference strain Pnstuer.38.95％ (37/95) of the isolates had the homologous locus sequences compared with the reference strain Ames Ancestor.The remaining strains were different from those completed sequenced strains.3 strains missed length of about 80 bp sequence in the PS-34 loci amplified gene fragment in which the tested SNP loci were included.9 strains were amplified negative at all SNP loci and Bacillus anthracis capsule plasmid genes were missing which was confirmed by capsule plasmid gene-speeific primers.Conclusion Results through analysis showed that single nucleotide genetic stability and specificity for capsule plasmid gene of Bacillus anthracis did exist in the Chinese isolates.The 6 discriminating SNP sites could be used as indicators in genotyping the Bacillus anthracis.