脱卤脱亚硫酸菌脱氯呼吸链的遗传分析

GENETIC ANALYSIS OF DESULFITOBACTERIUM DEHALOGENANS DECHL ORORESPIRATION CHAIN

以携有结合转座子Ｔｎ９１６的Ｅｎｔｅｒｏｃｏｃｃｕｓ ｆａｅｃａｌｉｓ ＪＨ２－２为供体，脱卤脱亚硫酸菌ＨＳＳ１（Ｄｅｓｕｌｆｉｔｏｂａｃｔｅｒｉｕｍ ｄｅｈａｌｏｇｅｎａｎｓ，Ｓｍ抗性突变株）为受体，在厌氧条件下，通过滤膜杂交、结合转移，将Ｔｈ９１６转移并插入到受体菌的染色体上，其转移频率为：１．１×１０－７～３×１０－８。 在丙酮酸／乳酸－３－氯－４－羟基苯氧乙酸、Ｔｃ、Ｓｍ培养基上，筛选脱氯呼吸的缺陷型突变株，并用反向ＰＣＲ（Ｉｎｖｅｒｓｅ－ＰＣＲ）技术，获得插入相应位点的染色体ＤＮＡ片段，进一步分析脱氯呼吸链的遗传背景。

Enteroccus faecilus JH2-2, carrying Tn916, wsa taken as donor, Desulfitobacterium dehalogenans asrecipient Under anaerobic condition , transfer Tn916 into the recipient by filter mating and conjugationg tranferring .The frequency was:1.1 ×10-7~3 ×10-6 per recipient. On the medium , containing Pyruvate / Lactate-CIOHPA,Tet. Sm, Screen mutants deficiency in dechororepiration. Obtain chromosome DNA fragment flanking Tn916, byInverse-PCR techniques,. then, investigate its genetic fundament involved dechlororespiration .