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大肠杆菌单拷贝rRNA操纵元菌株的构建及其生长特性研究

DOI: 10.7685/j.issn.1000-2030.2008.03.012, PP. 60-64

Keywords: 大肠杆菌,Red同源重组,rRNA操纵元,16SrRNA基因

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Abstract:

以1株染色体上的5个rRNA操纵元(rrnA、rrnD、rrnE、rrnG和rrnH)被敲除的大肠杆菌(Escherichiacoli)菌株SQ88为出发菌,运用λRed同源重组系统和卡那霉素抗性基因筛选重组子,并利用抗性基因两端的FRT位点通过位点专一性重组将抗性基因去除,最终成功构建了1株仅具有单拷贝rRNA操纵元(rrnB)的E.coli菌株――SQ88C。试验结果发现,此菌株并未出现明显的生长障碍,其生长速率和rRNA/蛋白值与出发菌株SQ88相比有所下降,但并不显著。从而证明了单拷贝rRNA操纵元在一定条件下能够满足大肠杆菌正常生长的需要,也为在大肠杆菌及其他菌株中快速精确地构建多rRNA基因缺失菌株提供了有益的参考,并可望在16SrRNA基因突变研究等方面发挥一定的作用。

References

[1]  萨姆布鲁克?J,拉塞尔 D W,黄培堂.分子克隆实验指南[M].北京:科学出版社.2002
[2]  Brunschede H,Dove T L,Bremer H.Establishment of exponential growth after a nutritional shift-up in Escherichia coli B/r:accumulation of deoxyribonucleic acid,ribonucleic acid,and protein[J].Journal of Bacteriology.1977,129(2):1020-1033
[3]  Condon C,French S,Squires C.Depletion of functional ribosomal RNA operons in Escherichia coli causes increased expression of the remaining intact copies[J].EMBO Journal.1993,12:4305-4315
[4]  Dammel C S,Noller H F.A cold-sensitive mutation in 16S rRNA provides evidence for helical switching in ribosome assembly[J].Genes and Development.1993,7:660-670
[5]  Ellwood M,Nomura M.Deletion of a ribosomal ribonucleic acid operon in Escherichia coli[J].Journal of Bacteriology.1980,143(2):1077-1080
[6]  Widom R L,Jarvis E D,Lafauci G.Instability of rRNA operons in Bacillus subtilis[J].Journal of Bacteriology.1988,170(2):605-610
[7]  Nomura M.Engineering of bacterial ribosomes:replacement of all seven Escherichia coli rRNA operons by a single plasmid-encoded operon[J].Proceedings of the National Academy of Sciences(USA).1999,96(5):1820-1822
[8]  Andersson S G E,Zomorodipour A,Winkler H H.Unusual organization of the rRNA genes in Rickettsia prowazekii[J].Journal of Bacteriology.1995,177(14):4171-4175
[9]  García-Marínez J,Acinas S G,Antón A I.Use of the 16S-23S ribosomal genes spacer region in studies of prokaryotic diversity[J].Journal of Microbiological Methods.1999,36:55-64
[10]  Asai T,Condon C,Voulgaris J.Construction and initial characterization of Escherichia coli strains with few or no intact chromosomal rRNA operons[J].Journal of Bacteriology.1999,181(12):3803-3809
[11]  Ellis H M,Yu D,Ditizio T.High efficiency mutagenesis,repair,and engineering of chromosomal DNA using single-stranded oligonucleotides[J].Proceedings of the National Academy of Sciences(USA).2001,98(12):6742-6746
[12]  Yu D,Ellis H M,Lee E C.An efficient recombination system for chromosome engineering in Escherichia coli[J].Proceedings of the National Academy of Sciences(USA).2000,97(11):5978-5983
[13]  Pace N R.A molecular view of microbial diversity and the biosphere[J].Science.1997,276:734-740
[14]  Condon C,Philips J,Fu Z Y.Comparison of the expression of the seven ribosomal RNA operons in Escherichia coli[J].EMBO Journal.1992,11:4175-4185
[15]  Woese C R.Bacterial evolution[J].Microbiological Reviews.1987,51(2):221-271
[16]  Osorio C R,Collins M D,Romalde J L.Variation in 16S-23S rRNA intergenic spacer regions in Photobacterium damselae:a mosaic-like structure[J].Applied and Environmental Microbiology.2005,71(2):636-645
[17]  Poteete A R.What makes the bacteriophage λ Red system useful for genetic engineering:molecular mechanism and biological function[J].FEMS Microbiology Letters.2001,201(1):9-14

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