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铜绿假单胞菌甘露醇阴性突变体中磷酸葡萄糖异构酶突变的表征及基因定位

Characterization and genetic mapping of phosphoglucoisomerase mutations in mannitol-negative mutants of Pseudomonas aeruginosa.

作者信息

Calligeros J E, Matsumoto H, Phibbs P V, Gates J E

机构信息

Department of Biology, Virginia Commonwealth University, Richmond, VA 23284, USA.

出版信息

Curr Microbiol. 1996 Dec;33(6):347-51. doi: 10.1007/s002849900126.

Abstract

Phosphoglucoisomerase (pgi) mutations in a number of independently isolated mannitol-negative mutants of Pseudomonas aeruginosa PAO1 were mapped on the chromosome by plasmid FP5-mediated conjugation and by cotransduction with the generalized transducing phages G101 and F116L. Mutant allele pgi-9001 exhibited linkage to ilvB, C-9059 (46-85%), car-9003 (93-100%), and pur-9047 (70%), but not with met-9011, in FP5-mediated conjugational crosses. All known pgi mutations and several previously uncharacterized mannitol-negative mutations exhibited transductional linkage to two independent car mutations at frequencies ranging from 13% to 42% and 53% to 99%, in transductional crosses mediated by phages G101 and F116L respectively. These pgi and mannitol-negative mutations also were cotransducible at very low frequencies (<1%) with two independent ilv mutations. Cotransduction of the car and ilv loci could not be detected. These data suggest the location of pgi within the first minute of the P. aeruginosa chromosome closely linked to the car marker and probably between the ilv and car loci. All of the mannitol-negative mutations that exhibited linkage to the car and ilv loci were characterized as pgi mutations by enzyme assays. A phenotypically similar, mannitol-negative mutant was shown to contain a mutation in glucose-6-phosphate dehydrogenase (zwf-9012) that maps to a different region on the chromosome.

摘要

通过质粒FP5介导的接合以及与通用转导噬菌体G101和F116L的共转导,对铜绿假单胞菌PAO1多个独立分离的甘露醇阴性突变体中的磷酸葡萄糖异构酶(pgi)突变进行了染色体定位。在FP5介导的接合杂交中,突变等位基因pgi - 9001与ilvB、C - 9059(46 - 85%)、car - 9003(93 - 100%)和pur - 9047(70%)连锁,但与met - 9011不连锁。在分别由噬菌体G101和F116L介导的转导杂交中,所有已知的pgi突变和几个先前未表征的甘露醇阴性突变与两个独立的car突变表现出转导连锁,频率分别为13%至42%和53%至99%..这些pgi和甘露醇阴性突变与两个独立的ilv突变也能以非常低的频率(<1%)共转导。未检测到car和ilv位点的共转导。这些数据表明pgi位于铜绿假单胞菌染色体第一分钟内,与car标记紧密连锁,可能位于ilv和car位点之间。所有与car和ilv位点连锁的甘露醇阴性突变通过酶分析被鉴定为pgi突变。一个表型相似的甘露醇阴性突变体被证明含有葡萄糖 - 6 - 磷酸脱氢酶(zwf - 9012)的突变,该突变位于染色体的不同区域。

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