Recombinant bovine resistin protein and preparation method thereof
A technology of resistin and protein, applied in the field of bioengineering, can solve the problems of protein conformation change, unfavorable activity research, oversized label, etc., and achieve the effect of high expression, simple operation and low cost
Inactive Publication Date: 2018-07-24
SICHUAN AGRI UNIV
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Problems solved by technology
[0004] The technical problem to be solved in the present invention is: the label contained in the protein expressed by the existing method for preparing bovine resistin protein is too large, which may easily cause changes in the protein conformation, which is not conducive to activity research and other issues
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[0028] The present invention will be described in detail below through specific embodiments in conjunction with the accompanying drawings. In the following examples, those that do not indicate specific experimental conditions are all according to conventional conditions well known to those skilled in the art, such as Sambrook J, Russell D.W., 2001, Molecular Cloning: A laboratory manual (3 rd ed), the conditions described in the Spring Harbor Laboratory Press, or as recommended by the manufacturer.
[0029] Example Preparation of recombinant bovine resistin protein by the method of the present invention
[0030] Material Remarks:
[0031] Pre-amplify the 347bp target gene, and insert the target gene fragment into the multiple cloning site of the pET-21a(+) expression vector. The construction method is a conventional method. Escherichia coli DH5a, BL21 and Rosetta were purchased from Tiangen Company.
[0032] The specific operation steps are as follows:
[0033] Amplificat...
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The invention belongs to the field of bioengineering, and particularly relates to a recombinant bovine resistin protein and a preparation method thereof. Aiming at the problems that a label containedin a protein expressed by the prior method for preparing the bovine resistin protein is too large, easily causes a change in the conformation of the protein, and is not conducive to an activity research and the like, the invention provides the preparation method of the recombinant bovine resistin protein, which comprises the following steps: amplifying a bovine resistin gene with a RT-PCR method;constructing a pET21a-RETN vector; transforming the pET21a-RETN vector into an escherichia coli; performing selective culture, PCR detection, double enzyme digestion, and gene sequencing; selecting plasmids with a target gene; transforming the pET21a-RETN vector into an escherichia coli BL21 and Rosetta; and inducing the expression of the bovine resistin protein. The method provided by the invention is simple in operation, low in cost, high in expression amount, and capable of preparing a large amount of the recombinant bovine resistin protein with high-efficiency, and lays a foundation for the research of physiological functions and application to the prevention and treatment of related diseases.
Description
technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a recombinant bovine resistin protein and a preparation method thereof. Background technique [0002] Resistin (Resistin, RETN) is a specific small molecular protein found in mouse adipose tissue when studying the mechanism of action of insulin sensitizer-thiazolidinediones derivatives (thiazolidiones). Current studies have found that it not only participates in lipid-glucose metabolism and mediates the energy level of the body; it even participates in the inflammatory response and is an important regulator of the process of inflammation. However, the lack of resistin raw materials hinders further research on its physiological functions and application to related diseases prevention and treatment. [0003] In the prior art, there are few researches on the preparation of bovine resistin by cloning and expression methods, and in previous studies, the cloning conditions we...
Claims
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Login to View More IPC IPC(8): C12N15/70C07K14/575
CPCC07K14/575C12N15/70
Inventor 左之才阳明贤岑垚马志宇姚彩霞沈留红邓俊良马晓平余树民苟丽萍
Owner SICHUAN AGRI UNIV