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Method for constructing cryptococcus albidus engineering bacterial strain for producing oleic acid in high yield

A technology of Cryptococcus albicans and engineering strains, applied in the field of genetic engineering, can solve the problems of low oil production rate, large distance, complex composition of oil components, etc., and achieve the effect of increasing oil production rate and increasing the content of oleic acid

Inactive Publication Date: 2010-11-24
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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Problems solved by technology

However, the current common oleaginous yeast has low oil production rate and complex oil composition, which still has a large distance from the raw material requirements for high-quality biodiesel production.

Method used

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  • Method for constructing cryptococcus albidus engineering bacterial strain for producing oleic acid in high yield

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Embodiment Construction

[0019] Below in conjunction with accompanying drawing and embodiment the present invention will be further described:

[0020] 1 Experimental materials

[0021] 1.1 Plant material

[0022] The developing fruits of Tunga paulica were collected from the Arboretum of the Subtropical Forestry Research Institute of the Chinese Academy of Forestry, the seeds were peeled off, quick-frozen in liquid nitrogen, and stored in a -80°C refrigerator for later use.

[0023] 1.2 Strains and plasmids

[0024] Cryptococcus albidus was purchased from the Culture Collection Center of the Institute of Microbiology, Chinese Academy of Sciences, and Escherichia coli DH5α, Agrobacterium tumefaciens EHA 105, LBA4404, and AGL-1 were preserved in our laboratory. The plasmid pCAMBIA2300 was preserved in our laboratory, and the cloning vector pGEM-T Easy was purchased from Promega.

[0025] 1.3 Main biochemical reagents

[0026] RNA extraction kits were purchased from Tianze Gene Engineering Co., Ltd....

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Abstract

The invention relates to a method for constructing a cryptococcus albidus engineering bacterial strain for producing oleic acid in high yield, and provides a yeast engineering bacterial strain for producing oleic acid in high yield. A vector uses pCAMBIA2300 as a basic framework, contains aleurites montana VeSAD genes, and can change an oil producing yeast fatty acid component so as to improve the oleic acid content. The method comprises the following steps of: (1) acquisition of a full-length sequence of a cDNA coding area of the enzyme cutting site-containing aleurites montana VeSAD gene; (2) construction of a VeSAD positive expression vector; (3) acquisition of a recombinant agrobacterium strain; and (4) acquisition of the cryptococcus albidus engineering bacterial strain. The method has the advantages that: the VeSAD genes separated from total RNA of aleurites montana seeds are constructed on the yeast expression vector pCAMBIA2300, and the VeSAD genes are guided into the cryptococcus albidus by adopting an agrobacterium-mediated method so that a synthetic gene of vegetable fatty acid is efficiently expressed in the yeast, the oil yield of the cryptococcus albidus is increased, and particularly the oleic acid content of the grease is improved.

Description

technical field [0001] The invention relates to the field of genetic engineering, and belongs to a method for constructing genetic engineering strains, in particular to a method for constructing an engineering strain of Cryptococcus albus with high oleic acid production. Background technique [0002] In response to the intensifying oil crisis, countries around the world are actively formulating and implementing new energy development strategies, hoping to find new oil substitutes through various possible channels, and biodiesel is one of them. At present, the raw materials for the production of biodiesel mainly depend on oil crops, woody oil plantations, animal fats and waste oils. Extracting oil from oil trees has a series of restrictive factors such as occupying arable land area, long cycle and high cost. The development of animal fat also faces a series of constraints. Therefore, it is of great significance to open up new production methods of plant-derived oils and fat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/81C12N1/19C12P7/64C12R1/645
Inventor 李纪元范正琪陈东亮田敏吴开云李辛雷
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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