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Cloning of mycobacterial histidine synthesis genes by complementation of a Mycobacterium smegmatis auxotroph
Author(s) -
Hinshelwood S.,
Stoker N. G.
Publication year - 1992
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1992.tb01468.x
Subject(s) - mycobacterium smegmatis , biology , complementation , cosmid , auxotrophy , operon , histidine , mutant , gene , cloning (programming) , genetics , escherichia coli , mycobacterium , mycobacterium tuberculosis , microbiology and biotechnology , bacteria , amino acid , medicine , tuberculosis , pathology , computer science , programming language
Summary Histidine‐requiring auxotrophs of Mycobacterium smegmatis were isolated following N ‐methyl‐ N′ ‐nitro‐ N ‐nitrosoguanidine treatment. One of these mutants, his 5, was transformed with an M. smegmatis shuttle cosmid library, and complementing clones were isolated at a frequency of approximately 1%. A 2.3 kb fragment was subcloned and sequenced, and found to contain the start of an operon including the hisD gene and part of the hisC gene. No hisG gene was detected upstream of hisD , suggesting that the regulation of histidine biosynthesis in mycobacteria may differ from that of Escherichia coli. The strategy used here will allow the molecular genetics of complex mycobacterial‐specific biosynthetic pathways involved in the virulence of pathogenic species to be studied.