检验医学
檢驗醫學
검험의학
LABORATORY MEDICINE
2014年
7期
750-754
,共5页
刘锦燕%倪培华%史册%魏冰%项明洁
劉錦燕%倪培華%史冊%魏冰%項明潔
류금연%예배화%사책%위빙%항명길
白念珠菌%氟康唑%ERG11 基因%错义突变%定点突变技术%酿酒酵母%异源性表达
白唸珠菌%氟康唑%ERG11 基因%錯義突變%定點突變技術%釀酒酵母%異源性錶達
백념주균%불강서%ERG11 기인%착의돌변%정점돌변기술%양주효모%이원성표체
Candida albicans%Fluconazole%ERG11 gene%Missense mutation%Site-directed mutagenesis%Saccharomyces cerevisiae%Heterologous expression
目的:研究临床耐氟康唑白念珠菌14-α脱甲基酶的 K143Q 氨基酸置换与氟康唑耐药形成的关系。方法运用体外定点突变技术构建 ERG11基因 A427C 突变型重组质粒,并利用酶切、连接构建 YES2/CT 酵母表达质粒。通过构建成的表达质粒在酿酒酵母中的异源性表达及体外药物敏感性表型的检测,分析其致 K143Q氨基酸置换与白念珠菌对氟康唑产生耐药的关系。结果真菌体外药物敏感性表型检测显示,含氨基酸置换的表达质粒转染于酿酒酵母 INVSc1后,氟康唑最低抑菌浓度(MIC)增加16倍。结论K143Q 可增强白念珠菌对氟康唑的耐药性。
目的:研究臨床耐氟康唑白唸珠菌14-α脫甲基酶的 K143Q 氨基痠置換與氟康唑耐藥形成的關繫。方法運用體外定點突變技術構建 ERG11基因 A427C 突變型重組質粒,併利用酶切、連接構建 YES2/CT 酵母錶達質粒。通過構建成的錶達質粒在釀酒酵母中的異源性錶達及體外藥物敏感性錶型的檢測,分析其緻 K143Q氨基痠置換與白唸珠菌對氟康唑產生耐藥的關繫。結果真菌體外藥物敏感性錶型檢測顯示,含氨基痠置換的錶達質粒轉染于釀酒酵母 INVSc1後,氟康唑最低抑菌濃度(MIC)增加16倍。結論K143Q 可增彊白唸珠菌對氟康唑的耐藥性。
목적:연구림상내불강서백념주균14-α탈갑기매적 K143Q 안기산치환여불강서내약형성적관계。방법운용체외정점돌변기술구건 ERG11기인 A427C 돌변형중조질립,병이용매절、련접구건 YES2/CT 효모표체질립。통과구건성적표체질립재양주효모중적이원성표체급체외약물민감성표형적검측,분석기치 K143Q안기산치환여백념주균대불강서산생내약적관계。결과진균체외약물민감성표형검측현시,함안기산치환적표체질립전염우양주효모 INVSc1후,불강서최저억균농도(MIC)증가16배。결론K143Q 가증강백념주균대불강서적내약성。
Objective To verify the correlation of K143Q amino acid substitution in 14α-demethylase and fluconzole resistance to Candida albicans.Methods The recombinant mutant plasmid with A427C in ERG11 gene was firstly constructed by site-directed mutagenesis,and YES2 /CT yeast expression plasmid was established by digestion and ligation technique.The heterologous expression in Saccharomyces cerevisiae and drug susceptibility in vitro were performed to analyze the correlation of K143Q amino acid substitution with resistance to fluconazole.Results The antifungal drug susceptibility test in vitro showed that the constructed plasmid with amino acid substitution transferred in Saccharomyces cerevisiae INVSc1 contributed to increase in fluconazole minimal inhibitory concentration (MIC)for 16 times.Conclusions The K143Q contributes to increase the resistance to fluconazole in Candida albicans clinical isolates.