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The Construction of Shuttle Vector between Zymomonas Mobilis and Escherichia Coli

Author: GaoWeiHua
Tutor: ZhangMinHua
School: Tianjin University
Course: Chemical processes
Keywords: Ethanol Zymomonas bacteria Shuttle plasmid
CLC: TQ920
Type: Master's thesis
Year: 2004
Downloads: 254
Quote: 5
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Abstract


The movement is a Gram-negative bacterium Zymomonas , micro - aerobic growth . Their sugar intake speed and fermentation alcohol speed are superior to conventional Saccharomyces cerevisiae , is believed to be great potential for development of ethanol -producing bacteria . Available only carbon source glucose, fructose and sucrose . This people a lot of work by genetic engineering to foreign gene into the bacteria , and thus broaden the range of available carbon source . This article describes the type of cloning in a Z.mobilis-E.coli shuttle plasmid . The isolated by Z.mobilis ATCC10988 2.7 kb plasmid pZM2 of SphI restriction sites into the E. coli plasmid pBR328 , building into a size of about 7.6 kb recombinant shuttle plasmid ( pZB1 ) . This plasmid having a the Apr Cmr resistance marker , ampicillin resistance gene inserted bit point of AhdI, PstI, PvuI , PSP , EcoRI in the chloramphenicol resistance gene inserted into the NcoI sites . The plasmid was successfully transformed into E. coli DH5a and Z. mobilis CP4 strain . At the same time , the stability of the plasmid in E. coli DH5a and Z. mobilis CP4 were studied. The Z.mobilis ATCC 10988 and CP4 growth in different media was investigated , and the results show that the T medium suitable for the growth of the test strains . Same time, examining the SDS- alkaline lysis method and Nester a plasmid extraction method , the identification of restriction endonuclease analysis and pZM2 .

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