Corynebacterium glutamicum genes encoding proteins involved in membrane synthesis and membrane transport

A coding and amino acid technology, applied in genetic engineering, plant genetic improvement, bacteria, etc., can solve time-consuming and difficult problems, and achieve the effect of optimizing activity, improving the ability of function, and enhancing vitality

Inactive Publication Date: 2007-12-05
BASF SE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, selecting strains that improve in the production of specific molecules is a time-consuming and difficult process

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0237] Example 1: Preparation of total genomic DNA of Corynebacterium glutamicum ATCC13032

[0238] The culture of Corynebacterium glutamicum (ATCC13032) was grown overnight in BHI medium (Difo) with shaking at 30°C. The cells were collected by centrifugation, the supernatant was discarded, and the cells were resuspended in 5 ml of buffer I (5% of the original volume of the culture-all specified volumes of bacteria have been calculated based on the 100 ml culture volume). The composition of buffer I: 140.34g / l sucrose, 2.46g / l MgSO 4 x 7H 2 O, 10ml / l KH 2 PO 4 Solution (100g / l, adjusted to pH 6.7 with KOH), 50ml / l M12 concentrate (10g / l(NH4) 2 SO 4 , 1g / l NaCl, 2g / l MgSO 4 x 7H 2 O, 0.2g / l CaCl 2 , 0.5g / l yeast extract (Difo), 10ml / l trace element mixture (200mg / lFeSO 4 x H 2 O, 10mg / l ZnSO 4 x 7 H 2 O, 3mg / l MnCl 2 x 4H 2 O, 30mg / lH3BO3, 20mg / l CoCl 2 x 6 H 2 O, 1mg / l NiCl 2 x 6H 2 O, 3mg / l Na 2 MoO 4 x 2H 2O, 500mg / l complexing agent (EDTA or citric acid), 100mg / l vitamin mixtur...

Embodiment 2

[0239] Example 2: Construction of a genomic library of Corynebacterium glutamicum ATCC13032 in E. coli

[0240] Using the DNA prepared as described in Example 1, according to a known clear method (see, for example, Sambrook, J. et al., (1989) "Molecular Cloning: A Laboratory Manual", Cold Spring Harbor Laboratory Press, or Ausubel, FM et al. (1994) ) "Current Protocols in Molecular Biology", John Wiley & Sons.) Construction of cosmid library and plasmid library.

[0241] Any plasmid or cosmid can be used. The specific plasmids used are plasmid pBR322 (Sutcliffe, JG (1 979) Proc. Natl. Acad. Sci. USA, 75: 3737-3741), pACYC177 (Change & Cohen (1978) J. Bacteriol 134: 1141-1156), pBS plasmid Series (pBSSK+, pBSSK-, etc.; Stratagene, LaJolla, USA) or cosmids such as SuperCosl (Stratagene, LaJolla, USA) or Lorist6 (Gibson, TJ, Rosenthal A. and Waterson, RH (1987) Gene 53:283-286). The plasmid pSL109 can be used to construct a gene library specifically for Corynebacterium glutamicum (Le...

Embodiment 3

[0242] Example 3: DNA sequencing and computational function analysis

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PUM

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Abstract

Isolated nucleic acid molecules, designated MCT nucleic acid molecules, which encode novel MCT proteins from Corynebacterium glutamicum are described. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing MCT nucleic acid molecules, and host cells into which the expression vectors have been introduced. The invention still further provides isolated MCT proteins, mutated MCT proteins, fusion proteins, antigenic peptides and methods for the improvement of production of a desired compound from C. glutamicum based on genetic engineering of MCT genes in this organism.

Description

[0001] This application is a Chinese patent application entitled "Corynebacterium glutamicum gene encoding stress, resistance and tolerance proteins" with the application date of June 23, 2002 (national application number No.00811817.5, international The application number is PCT / IB00 / 00922) divisional application. Technical field [0002] Related application [0003] This application requires the U.S. provisional patent application serial number 60 / 141031 filed on June 25, 1999, the U.S. patent provisional application serial number 60 / 142692 filed on July 1, 1999 and the U.S. provisional patent application filed on August 27, 1999 Priority of serial number 60 / 151214. The application also claims priority for the following patent applications: German patent application number 19930429.7 filed on July 1, 1999, German patent application number 19931413.6 filed on July 8, 1999, German patent application filed on July 8, 1999 No. 19931457.8, German patent application number 19931541.8...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C07K14/34C12N1/21C12P1/04C12Q1/68G01N33/68C07KC12NC12PC12Q
Inventor M·波姆佩朱斯B·克雷格尔H·施雷德尔O·策尔德G·哈贝豪尔H·－S·李H·－J·李
Owner BASF SE
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