Please wait a minute...

中国生物工程杂志

China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (02): 50-55    DOI:
    
Location of Some Genes Related to One Carbon Metabolism in Methylobacterium sp. MB200 and Cloning of mtdA and mtdB Genes
SONG Xiu-peng1,2,3, WU Bo1,2,3, SHEN Pei-hong1,2,3, JIANG Cheng-jian1,2,3, TIAN Dan-dan1,2,3, TANG Xian-lai1,4
1. The College of Iife Science and Technology, Guangxi University, Nanning 530005, China;
2. Key laboratory of Microbial and Plant Genetic Engineering of Ministry of Education, Nanning 530005, China;
3. Key Laboratory of Subtropical Bioresource Conservation and Utilization of Guangxi, Nanning 530005, China;
4. The Science and Technology Department of Guangxi, Nanning 530005, China
Download: HTML   PDF(1451KB) HTML
Export: BibTeX | EndNote (RIS)      

Abstract  

The partial mutants library of Methylobacterium sp. MB200 was constructed by using plasmid pTnMod-RKm’ and 11554 mutants were obtained which could survive in MMII containing two antibiotics (Nm and Km). Rescreening results showed that 333 strains could not use methanol as the sole carbon source,these strains were preliminarily considered that their one carbon metabolism way had been destroyed. This laid the foundation for cloning genes related to one carbon metabolism. The location analysis was carried out by TAIL-PCR, based on the characters of pTnMod-RKm’ and the full sequence of mtdA and mtdB genes were cloned .



Key wordsMethylobacterium      Mutants library pTnMod-RKm’      TAIL-PCR      mtdA      mtdB     
Received: 13 October 2010      Published: 18 February 2011
ZTFLH:  Q785  
Cite this article:

SONG Xiu-peng, WU Bo, SHEN Pei-hong, JIANG Cheng-jian, TIAN Dan-dan, TANG Xian-lai. Location of Some Genes Related to One Carbon Metabolism in Methylobacterium sp. MB200 and Cloning of mtdA and mtdB Genes. China Biotechnology, 2011, 31(02): 50-55.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I02/50

[1] Tanson R S, Hanson T E. Methanutrophic hacteria. Microbiol Rev,1996,60(2): 439-471.
[2] Chisteserdova L, Chen S W, Lapidus A, et a1. Methylotrophy in Methylobacterium extorquens AM1 from a genomic point of view. J Bacteriol, 2003, 185(10): 2980-2987.
[3] Shen P H. Wu B. Over-expression of a hydroxypyvavate reductase in Methylobacterium MB200 enhances glyoxylate accumulation. J Ind Microbiol Biotehnol, 2007, 34(10): 657-663.
[4] Shen P H, Chao H J, Jiang C J, et a1. Enhancing production of L-serine by increasing the glyA gene expression in Methylobacterium sp. MB200. Appl Biochem Biotechnol, 2009,160(3):740-750.
[5] Vandien S J, Marx C J, O’Brlen B N, et a1. Genetic characterization of the carotenoid biosynthetic pathway in Methylobacterium extorquens AM1 and isolation of a colorless mutant.Appl Environ Microbiol, 2003, 69(12): 7563-7566.
[6] Marx C J, O’Bfien B N, Breezce J, et a1. Novelmethylotrophy genes of Methylobacterium extorquens AM1 identified by using transposon mutagenesis including a putative dihydromethanopterin reductase. J Bacteriol, 2003, 185(2): 669-673.
[7] Dennis J J, Zylstra G J. Plasposons: modular self-cloning minitransposon derivatives for rapid genetic analysis of gram-negative bacterial genomes. Appl Environ Microbiol, 1998,64(7): 2710-2715.
[8] Kalyuzhnaya M G, Lidstrom M E. QscR——mediated transcriptional activation of serine cycle genes in Methylobacterium extorquens AM1. J Bacteriol, 2005, 187(21): 7511-7517.
[9] Hanahan D. Studies on transformation of Escherichia coli with plasmid, J Mol Bid, 1983, 166: 557.
[10] Figurski D, Helinski D R.Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans.PNAS, 1979, 76:1648-1652.
[11] Green P N. Methylobacterium. Protebacteria: Alpha and Beta Subclasses. New York:Springer New York,2006.257-265.
[12] Liu Y G, Whittier R F. Thermal asymmetric interlaced PCR:automatable amplification and sequencing of insert end flagments from P1 and YAC clones for chromosomal walking. Genomics,1995, 25(3):674-681.
[13] Liu Y G, Chen Y, Zhang Q. Amplification of genomic sequences flanking T-DNA insertions by thermal asymmetric interlaced polymerase chain reaction. Methods Mol Biol, 2005, 286:341-348.
[14] Hong D F, Wan L L, Yang G S, The evaluation of the Flanking sequence’s cloning method. Molecular Plant Breeding. 2006, 4(2): 280-288.
[15] 罗丽娟, 施季森, 一种DNA侧翼序列分离技术——TAIL-PCR. 南京林业大学学报(自然科学版),2003,27(4): 87-90. Luo L J, Shi J S.Journal of Nanjing Forestry University(nature science edition),2003,27(4): 87-90.
[16] Crowther G J, Kosaly G. Formate as the main branch point for methylotrophic metabolism in Methylobacterium extorquens AM1, J Bacteriol, 2008, 190(14): 5057-5062.