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Construction of Lescpth5 bait vector and Detection of Its Self-activated Activity

Plant Diseases and Pests 2011年第4期2卷 1-2,5页

作者：张月娟万一陈华民张琨赵廷昌陕西省微生物研究所陕西西安710043 中国农业科学院植物保护研究所北京100193

[ Objective ] The paper was to construct bait vector pGBKTT-Lescpth5, and detect its self-activated transcription activity and the toxicity on yeast cells. [ Method] Lescpth5 was amplified by PCR technique, and connected into bait vector pGBKTT. The recombined vector was transformed into yeast competent ceils AH109 and carried out self-activated detection and toxicity detection in the *** medium. [ Result] Digestion and sequencing results showed that bait vector pGBKT7-Lescpth5 was suecessfully constructed with correct reading frame. Self-activated activity and toxicity detection results showed that bait vector had no self- activated activity on yeast strain AH109, which also had no toxicity on yeast. [Condusion] bait vector successfully constructed could be used in yeast two-hybrid system, which laid the foundation for screening of cDNA library in the next step.

关键词： Pseudomonas syringae pv. tomato bait vector Yeast two-hybrid Self-activated China

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The Self-activated Experimental of T_(1083) Substitution Mutation vector pGBKT7-TS in Yeast

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Agricultural Science & Technology 2010年第3期11卷 65-67页

作者：袁亮纪耀坤张伟彬商丘职业技术学院河南商丘476100

[Objective] The research aimed to study the self-activation test of inverting T1083 substitution mutation BD fusion vector pGBKT7-TS into yeast,and discuss whether its expression product can be used as bait for further two-hybrid screening.[Method] T1083 substitution mutation BD fusion vector pGBKT7-TS was inverted into yeast to make the self-activation and protein expression toxic detection test.[Result] This expression product of the fragment was inactive and had no toxic to yeast *** could be used as bait for further two-hybrid screening.[Conclusion] The research laid the experimental foundation for further study of the effect of T1083 substitution mutation on the interaction of NtKrp and its target partner.

关键词： pGBKT7-TS vector Yeast two-hybrid bait vector Self-activation

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