摘要
为研究猪链球菌细菌素基因能否在同种和不同种属之间转移,本研究对供体菌xj-144的红霉素耐药基因ermB和四环素耐药基因tetO进行PCR鉴定,然后以携带猪链球菌细菌素基因的菌株xj-144为供体菌,分别以猪链球菌BAA和粪肠球菌JH2-2株为受体菌进行膜接合转移试验,并对接合子进行PCR检测,对经检测正确的接合子进行药敏试验,同时对接合子的细菌素基因簇和供体菌的可移动遗传元件经PCR鉴定,并对整合性接合元件进行基因序列分析。结果显示,供体菌xj-144携带红霉素耐药基因ermB和四环素耐药基因tetO,与受体菌BAA和JH2-2株分别进行滤膜接合转移试验后正确获得了接合子BM-1和JM-1,接合效率分别为8×10;和8×10^(-7)。药敏试验结果显示,供体菌xj-144对红霉素和四环素耐药,受体菌BAA和JH2-2株对红霉素和四环素均敏感,接合子BM-1和JM-1均对红霉素和四环素耐药。且接合子经PCR均扩增出了猪链球菌细菌素的suicin65基因簇;同时,从供体菌和接合子中均经PCR扩增到转座子整合酶int基因;序列分析结果显示,细菌素基因簇与耐药基因ermB、tetO共定位于一个新的整合性接合元件ICESsuxj-144中。上述结果首次证实了猪链球菌细菌素基因可以跨种属水平转移,并且该基因的水平转移与转座子有密切关系,本研究为进一步探究细菌素基因的转移提供了参考依据。
To investigate whether Streptococcus suis bacterin gene can be transferred between the same species and different species,the erythromycin resistance gene ermB and tetracycline resistance gene tetO in donor strain xj-144 were identified by PCR in this study.The membrane conjugation test was carried out using the xj-144 as donor bacteria and the S.suis BAA and E.faecalis JH2-2 as recipient bacteria,respectively.The resulting zygotes were identified by PCR.Test the correct zygotes for drug sensitivity The bacteriocin gene cluster in the positive zygotes and the transferable genetic elements of the donor bacteria were verified by PCR.Analysis of integrated and conjugative elements for gene sequence.The results showed that the donor strain xj-144 contained ermB gene and tetO gene,and the positive zygotes BM-1 and JM-1 were successfully obtained by filtration membrane conjugation test with recipient strain BAA and JH2-2,and the conjugation efficiency of 8×10;and 8×10;,respectively.By the drug sensitivity test,the donor strain xj-144 was resistant to erythromycin and tetracycline,the recipient bacteria BAA and JH2-2 were sensitive to erythromycin and tetracycline,and zygotes BM-1 and JM-1 were resistant to erythromycin and tetracycline.The gene cluster encoding S.suis bacteriocin was identified by PCR in zygotes.Meanwhile,transposon integrase gene int was amplified by PCR from both the donor and the zygote.Sequence analysis showed that the bacteriocin gene cluster and drug resistance genes ermB and tetO co-positioning with a new ICESsuxj-144.This study confirms for the first time that S.suis bacteriocin gene can be transmitted horizontally,and the horizontal transfer of this gene is closely related to transposons,which provides a basis for further research on bacteriocin gene transfer.
作者
王凌利
杨亲
祝瑶
杨文琳
曹俊
刘思国
张万江
WANG Ling-li;YANG Qin;ZHU Yao;YANG Wen-lin;CAO Jun;LIU Si-guo;ZHANG Wan-jiang(State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research Institute,Chinese Academy of Agriculture Science,Harbin 150069,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2022年第5期465-470,507,共7页
Chinese Journal of Preventive Veterinary Medicine
基金
黑龙江省自然科学基金优秀青年项目(YQ2019C031)
中国博士后科学基金(2017M620076)。
关键词
猪链球菌细菌素
接合转移
基因水平转移
转座子
Streptococcus suis bacteriocin
conjugal transfer
horizontal gene transfer
transposon
