生物技术通讯
生物技術通訊
생물기술통신
LETTERS IN BIOTECHNOLOGY
2014年
4期
504-507
,共4页
蔡红艳%陈保立%梁未丽%王多春%李杰%段国贤%阚飙
蔡紅豔%陳保立%樑未麗%王多春%李傑%段國賢%闞飆
채홍염%진보립%량미려%왕다춘%리걸%단국현%감표
环腺苷酸受体蛋白%高丝氨酸脱氢酶%天冬氨酸%霍乱弧菌
環腺苷痠受體蛋白%高絲氨痠脫氫酶%天鼕氨痠%霍亂弧菌
배선감산수체단백%고사안산탈경매%천동안산%곽란호균
cAMP receptor protein%homoserine dehydrogenase%aspartate%Vibrio cholerae
目的:研究霍乱弧菌中环腺苷酸受体蛋白(CRP)对高丝氨酸脱氢酶(HDH)是否具有调控作用。方法:通过搭桥PCR将HDH基因启动子中CRP潜在结合位点替换为CRP保守结合位点,分别将替换前后的启动子序列导入报告质粒,转化霍乱弧菌野生株C7258及CRP缺失株WL7258,比较荧光值的差异。结果:改造前报告质粒在野生株中的荧光值高于CRP缺失株,改造后报告质粒的荧光值在2株菌中均有升高,但在野生株中升高更明显。结论:CRP调控HDH的转录,但不是惟一调控因素。
目的:研究霍亂弧菌中環腺苷痠受體蛋白(CRP)對高絲氨痠脫氫酶(HDH)是否具有調控作用。方法:通過搭橋PCR將HDH基因啟動子中CRP潛在結閤位點替換為CRP保守結閤位點,分彆將替換前後的啟動子序列導入報告質粒,轉化霍亂弧菌野生株C7258及CRP缺失株WL7258,比較熒光值的差異。結果:改造前報告質粒在野生株中的熒光值高于CRP缺失株,改造後報告質粒的熒光值在2株菌中均有升高,但在野生株中升高更明顯。結論:CRP調控HDH的轉錄,但不是惟一調控因素。
목적:연구곽란호균중배선감산수체단백(CRP)대고사안산탈경매(HDH)시부구유조공작용。방법:통과탑교PCR장HDH기인계동자중CRP잠재결합위점체환위CRP보수결합위점,분별장체환전후적계동자서렬도입보고질립,전화곽란호균야생주C7258급CRP결실주WL7258,비교형광치적차이。결과:개조전보고질립재야생주중적형광치고우CRP결실주,개조후보고질립적형광치재2주균중균유승고,단재야생주중승고경명현。결론:CRP조공HDH적전록,단불시유일조공인소。
Objective: To elucidate the effect of cAMP receptor protein(CRP) on the transcription of homoserine dehydrogenase(HDH) in Vibrio cholerae. Methods: Using overlap PCR, the possible CRP bingding site in the pro-moter of HDH gene was replaced by consensus sequence. The transcriptional reporter plasmids containing the origi-nal promoter or changed promoter were constructed, and these plasmids were introduced into the V.cholerae C7258 and CRP mutant strain WL7258 by conjugation. The transcription level was compared by monitoring luminescence value in C7258 and WL7258. Results: Our data showed that luminescence value of plasmid which containing the original promoter was higher in C7258 than WL7258. After the promoter was changed, luminescence value of plas-mid were raised in two strains, especially in C7258. Conclusion: The results demonstrated that CRP regulates the transcription of HDH in V.cholerae, but CRP was not the only regulation factor.