23 Jul Pichia pastoris engineering bacteria of secretory expression human III type alpha chain collagen and construction method and application thereof
Abstract
The invention relates to a pichia pastoris engineering bacteria of secretory expression human III type alpha chain collagen and a construction method and application thereof. The type III collagen can be used as a biomedical material, but large-scale production is not realized. According to the invention, RNA is extracted from human placenta tissues, a cDNA library is built through inverse transcription, then mature peptide chain gene of type III alpha chain collagen is amplified by using a PCR (Polymerase Chain Reaction) method, the the gene is connected with a pichia pastoris expression vector pPIC9K, and plasmid pPIC9K-CO3A1 is structured; after linearization, the well structured plasmid pPIC9K-CO3A1 is led into pichia pastoris SMD1168, and a high copy pichia pastoris engineering bacteria SMD1168-CO3A1 is obtained through resistance screening. The engineering bacteria provided by the invention can effectively and safely secrete human III type alpha chain collagen, has higher biological activity, the collagen can be mixed with other inorganic materials to serve as a biological and degradable biomedical material, and used for tissue defects, repair and the like.
Year: 2014
Country: CN
Doc No: 103725623