Transgenic pichia pastoris gene engineering bacteria and construction method and application thereof

2013 
The invention relates to transgenic pichia pastoris gene engineering bacteria and a construction method and application thereof. I-type collagen can be applied to a biomedical material, but large-scale industrial production cannot be realized. The construction method for the transgenic pichia pastoris gene engineering bacteria comprises the steps of extracting a main RNA (ribonucleic acid) from a human placenta tissue, performing reverse transcription to obtain cDNA (deoxyribonucleic acid), and cloning an I-type collagen alpha1 chain gene by taking the cDNA as a template; modifying the I-type collagen alpha1 chain gene, and connecting the I-type collagen alpha1 chain gene with an expression carrier pPIC9K to construct a plasmid pPIC9K-Col1a1; performing enzyme linearization on the constructed plasmid pPIC9K-Col1a1, and introducing pichia pastoris SMD1168; converting and putting the human I-type collagen alpha1 chain gene into the pichia pastoris SMD1168 to obtain the transgenic pichia pastoris gene engineering bacteria. The engineering bacteria disclosed by the invention can efficiently and safely secrete an I-type collagen alpha1 chain protein outside cells and is higher in bioactivity; the protein can be mixed with other inorganic materials and is used as a biodegradable biomedical material for tissue defection, restoration and the like.
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