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[菊欧文氏菌果胶酸裂解酶基因在大肠杆菌细胞中的克隆]

[Cloning of pectate-lyase genes of Erwinia chrysanthemi in Escherichia coli cells].

作者信息

Evtushenkov A N, Shevchik V E, Fomichev Iu K

出版信息

Mol Gen Mikrobiol Virusol. 1986 Apr(4):19-24.

PMID:3025699
Abstract

Erwinia chrysanthemi DNA fragment digested by restriction endonuclease EcoRI and carrying the gene EC16 determining the synthesis of pectatelyase with Rf 0.20 and mol. mass 40kD has been cloned in plasmid pUC 9 plasmid in Escherichia coli HB101 cells. Three genes for pectatelyases of Erwinia chrysanthemi ENA49 have been cloned in vector phage lambda 47.1 in Escherichia coli cells. Two genes determining the synthesis of pectatelyases with Rf 0.06 and 0.19 and mol. masses 40 kD and 39 kD have been cloned as a part of an 7 kb Eco RI-fragment, that suggested their close location on the chromosome of Erwinia chrysanthemi ENA49. All of the cloned pectatelyase genes are expressed constitutively with pectatelyases accumulating in periplasm and being unable to secret into the cultural medium.

摘要

经限制性内切酶EcoRI消化的、携带决定果胶酸裂解酶合成的基因EC16(Rf为0.20,分子量为40kD)的菊欧文氏菌DNA片段已克隆到大肠杆菌HB101细胞的质粒pUC 9中。菊欧文氏菌ENA49的三个果胶酸裂解酶基因已克隆到大肠杆菌细胞的载体噬菌体λ47.1中。决定Rf为0.06和0.19、分子量为40kD和39kD的果胶酸裂解酶合成的两个基因已作为一个7kb Eco RI片段的一部分被克隆,这表明它们在菊欧文氏菌ENA49染色体上的位置相近。所有克隆的果胶酸裂解酶基因都是组成型表达的,果胶酸裂解酶积累在周质中,无法分泌到培养基中。

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