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欧文氏菌果胶酸裂解酶基因中的Mu - lac插入定向诱变

Mu-lac insertion-directed mutagenesis in a pectate lyase gene of Erwinia chrysanthemi.

作者信息

Diolez A, Coleno A

出版信息

J Bacteriol. 1985 Sep;163(3):913-7. doi: 10.1128/jb.163.3.913-917.1985.

DOI:10.1128/jb.163.3.913-917.1985
PMID:2993251
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC219219/
Abstract

The pelC gene, which encodes one of the five major pectate lyase (PL) isoenzymes in Erwinia chrysanthemi 3937, designated PLc, was subcloned from a hybrid lambda phage into a pBR322 derivative and mutagenized with a mini-Mu-lacZ transposable element able to form fusions to the lacZ gene. One plasmid (pAD1) which had an inactivated pelC gene and a Lac+ phenotype was selected in Escherichia coli. This plasmid was introduced into Erwinia chrysanthemi, and the pelC::mini-Mu insertion was substituted for the chromosomal allele by homologous recombination. This strain lacks the PLc isoenzyme. This Erwinia chrysanthemi strain has a Lac+ phenotype that is inducible by polygalacturonate, as are the wild-type PL activities.

摘要

pelC基因编码菊欧文氏菌3937中五种主要果胶酸裂解酶（PL）同工酶之一，命名为PLc，该基因从杂交λ噬菌体亚克隆到pBR322衍生物中，并用能够与lacZ基因形成融合体的mini-Mu-lacZ转座元件进行诱变。在大肠杆菌中筛选出一个具有失活pelC基因和Lac+表型的质粒（pAD1）。将该质粒导入菊欧文氏菌，通过同源重组，pelC::mini-Mu插入取代了染色体等位基因。该菌株缺乏PLc同工酶。该菊欧文氏菌菌株具有Lac+表型，该表型可被聚半乳糖醛酸诱导，野生型PL活性也如此。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32f/219219/e0566b996a2b/jbacter00220-0110-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32f/219219/ffd5cd54083b/jbacter00220-0108-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32f/219219/6847cc0cfe48/jbacter00220-0109-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32f/219219/1ae8f964d175/jbacter00220-0110-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32f/219219/e0566b996a2b/jbacter00220-0110-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32f/219219/ffd5cd54083b/jbacter00220-0108-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32f/219219/6847cc0cfe48/jbacter00220-0109-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32f/219219/1ae8f964d175/jbacter00220-0110-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32f/219219/e0566b996a2b/jbacter00220-0110-b.jpg

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