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Chick pea kluyveromyces expression system and its use

A Kluyveromyces and expression system technology, applied in the field of Kluyveromyces chickpea expression system, can solve problems such as difficulties, unclear genetic background of strains, lack of selectable markers, etc.

Inactive Publication Date: 2003-04-09
FUDAN UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

But utilize Kluyveromyces chickpea to construct transformation system but do not see report, this mainly has following several reasons: 1, because the genetic background of this bacterial strain is very unclear, so utilize this bacterial strain to carry out the work of genetic manipulation seldom, This caused great difficulty
2. There is no host bacteria that can be used for the transformation system, because there is little scientific research work using this strain as a material, so no one has constructed a host bacteria that can be used for transformation
3. Lack of ideal carrier
4. Lack of effective selectable markers
5 No transformation method suitable for this strain has been established

Method used

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  • Chick pea kluyveromyces expression system and its use
  • Chick pea kluyveromyces expression system and its use
  • Chick pea kluyveromyces expression system and its use

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Embodiment Construction

[0030] The present invention is further described below by way of examples.

[0031] 1. Cloning of URA3 gene:

[0032] A yeast-large intestine shuttle plasmid pRS424 was used to construct the gene library of the Y179 strain, and the gene library was transformed into Saccharomyces cerevisiae strain W303-1B. By speculating that the Y179 URA3 gene and the ura3 of the Saccharomyces cerevisiae strain can be functionally complementary, the Y179 strain was screened. URA3 gene (SEQ.1). The gene encodes orotidine-5'-phosphate decarboxylase, which has 801 coding regions and a total of 267 amino acids. The nucleotide sequence homology with Saccharomyces cerevisiae is 80%, and the amino acid homology is 87%.

[0033] 2. Construction of Y179ura-strain

[0034]We used modern molecular genetics methods to mutate the typical strain of Kluyveromyces chickpea (Kluyveromycescicerispours Y179), and obtained the mutant strain of Kluyveromyces chickpea Y179ura- (preservation number is CCTCC NO: ...

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Abstract

The present invention provides auxotrophic chick pea Kluyveromyces capable of being used in gene engineering host bacillus, polynucleotides carrier constituted with the URA3 of the strain, chemical and electric transforming method to transform foreign gene into host bacillus, and the obtained gene engineering chick pea Kluyveromyces strain with specific foreign gene.

Description

field of invention [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to a Kluyveromyces chickpea expression system and its application, including a vector system, a host strain, a transformation method and the like required for the construction of genetic engineering strains. Background technique [0002] Due to the rapid development of molecular biology in recent years, the high-efficiency expression of foreign genes has become a very important field in applied research and basic theoretical research. A good expression system (Hashimoto, Yoshio; Miki, Takeyoshi; Mukae, Masami; Ueda, Tadashi; Imoto, Taiji. 1998 Construction of a yeast expression system with positive selection for gene insertion in the absence of a specific phenotype. GENE 207 (2 ): 167-170) At first there needs to be a transformation system (Kondo, K., Saito, T., Kajiwara, S., Takagi, M. and Misawa, N.A Transformation system for the yeast Candida utilis) sui...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/12C12N15/81
Inventor 李育阳张晋徐帆洪文铁桥邸玉君霍克克
Owner FUDAN UNIV
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