Eukaryotic expression method of molt-inhibiting hormone-1 gene of mitten crab

Abstract

The present invention discloses a eukaryotic expression method of molt-inhibiting hormone-1 gene of mitten crab, including the steps of constructing a eukaryotic expression vector of the molt-inhibiting hormone-1 gene of the mitten crab and eukaryotic expressing molt-inhibiting hormone-1 gene of the mitten crab. The eukaryotic expression method successfully expresses an MIH protein; provides a theoretical basis for further understanding the internal mechanism of the precocious crab gonad early development, understanding the function mechanism of the crab molt gland, and further establishing technical measures to control crab precocious puberty; and obtains artificial regulation basis of achieving growing development and reproduction.

Description

technical field [0001] The invention relates to a gene expression method, in particular to a eukaryotic expression method of the Chinese mitten crab molting inhibitory hormone 1 gene. Background technique [0002] Moulting is a symbolic feature of the growth and development of crustaceans. It runs through the individual development of crustaceans and is regulated by both the nervous system and the endocrine system. This regulatory process is manifested as two antagonistic hormone actions. Stimulation of ecdysone and inhibition of M IH. The content of MIH produced in crab eyestalk is very small, and the similarity in size and primary structure of other neuropeptides in the family limits the isolation and purification of MIH protein directly from animals. Therefore, prokaryotic and eukaryotic protein expression systems are often used to produce relatively large quantities of recombinant MIH (Yodmuang et al., 2004; Lee and Watson, 2002; Ohira et al., 1999; Sun, 1997). Song X...

AI Technical Summary

Problems solved by technology

However, when expressing some proteins, many difficulties will arise

A large amount of exogenous protein expressed in E.coli exists in the cell in the form of insoluble inclusion bodies, and the separation and fragmentation of inclusion bodies will usually cause the inactivation of the target protein; the expression of eukaryotic proteins in E.coli will have More problems: Since eukaryotic genes usually contain introns, correct splicing and splicing cannot be done in E.coli, so its cDNA sequence must be expressed

In addition, many proteins in eukaryotes are glycoproteins. Using E.coli as an expression host cannot perform correct post-translational processing such as glycosylation of eukaryotic proteins, and it is difficult to obtain an active eukaryotic expression system

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