Preparation and application of recombinant porcine epidermal growth factor
A technology of epidermal growth factor and gene, applied in the field of genetic engineering, to avoid drug resistance and environmental pollution, improve economic benefits, and promote intestinal epithelial cell proliferation
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Embodiment 1
[0040] Embodiment 1: prepare the preparation of novel active protein as follows:
[0041] 1.1 Construction of recombinant porcine EGF expression plasmid
[0042] The relevant gene and nucleotide sequence of recombinant porcine epidermal growth factor (SEQ ID NO.1) were obtained through GenBank query, and E. 28a-rpEGF and pET-28a-SUMO-rpEGF expression plasmids.
[0043] 1.2 Induced expression of recombinant porcine EGF engineering bacteria
[0044] The constructed expression plasmids pET28a-rpEGF, pET32a-rpEGF and pET28a-SUMO-rpEGF were respectively transferred into competent cells prepared by BL21 / DE3.0. : 100 (for example, 2ml strains are inoculated into 100ml medium) inoculated in LB liquid medium containing resistance, cultured at 37°C with shaking at 220r / min until OD 600 When it is 0.5 to 0.6, add IPTG (to make the final concentration 0.5mmol / L) to induce expression for about 5 hours, take 1ml of the bacterial liquid induced to express, centrifuge at 12000r / min at 4°C ...
Embodiment 2
[0049] Embodiment 2: the application of novel active protein in feed additive:
[0050] EGF involves the regulation of a large number of ion channels and transport proteins, and promotes the absorption of nutrients in the intestine, such as regulating the absorption of phosphorus in the small intestine, maintaining normal cell functions, and promoting the absorption of glutamine by the epithelium. The special structure of EGF makes it highly resistant to heat and acid, and can resist digestion by trypsin and pepsin, which determines that it can act on the gastrointestinal tract for a long time. After oral administration of EGF to young piglets, it still retains its biological activity in the intestinal tract, promoting the growth and maturation of the gastrointestinal tract after birth.
[0051] Relevant literature shows that pEGF is beneficial to the early embryonic development of pigs, increasing the litter size of sows, improving the survival rate of piglets and promoting t...
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