Temperature-sensitive polypeptide and coding gene thereof and preparation method and application thereof
An encoding gene and temperature-sensitive technology, applied in the field of temperature-sensitive polypeptides and their encoded genes, can solve problems such as poor synthesis and difficulty in obtaining temperature-sensitive polypeptides, and achieve the effects of high recovery efficiency, simple and fast development, and wide application range.
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[0038] The principles and features of the present invention are described below in conjunction with the accompanying drawings, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.
[0039] 1. Materials and Reagents
[0040] Restriction endonuclease, T4 polynucleotide kinase, calf intestinal alkaline phosphatase, and T4 DNA ligase were all purchased from NEB Company. DNA Marker was purchased from Nearshore Protein Technology Co., Ltd., and pre-stained protein Marker was purchased from Thermo Scientific. The plasmid extraction kit was purchased from OMEGA BIO-TEK, and the gel recovery kit was purchased from Beijing Broadtech Bio-Gene Technology Co., Ltd. The expression plasmid pET-28a, strain E.coli Top10 and BL21(DE3) are preserved by our laboratory.
[0041] 2. Design the monomeric gene of thermosensitive polypeptide
[0042] In the course of our research, we found that the polypeptide compo...
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