Method for scale selection of microorganism alkaliproof related genes

Abstract

The invention discloses a method to sieve scale microbe alkali proof relative gene in microbe engineering domain, which is characterized by the following: collecting typical basic environment soil sample; rough-extracting; purifying; getting macro-genom DNA of soil sample; electrization-inverting bacillus coli defect strain E with connecting product between macro-genom DNA part enzyme cutting fragment and carrier; sieving; getting positive colony; proceeding sub-clone and functional appraise for alkali proof related gene of positive clone. This invention can be used to breeding of alkali proof and molecule of microbe.

Description

Technical field:

[0001] The invention belongs to the field of microbial engineering and relates to a method for large-scale screening of genes related to microbial alkali resistance. Background technique:

[0002] Internationally, Escherichia coli defective strain E is used to clone salt-tolerance-related genes from microorganisms. The method is: extract the DNA of the purely cultured microorganism, partially digest it with a restriction endonuclease, connect the digested fragment to an appropriate vector, and use the ligated product to transform E. coli defective strain E, and then, in a certain concentration The transformants were screened on the solid medium of NaCl or LiCl, so as to obtain the recombinants carrying the genes related to salt tolerance. In fact, it is also possible to use E. coli defective strain E to clone genes related to alkali resistance. Alkali resistance-related genes refer to genes related to intracellular pH homeostasis. But so far, no one has u...