Pig intestinal epithelial cell promoter SIECP and application thereof
A promoter and molecular technology, applied in the biological field, can solve problems such as the cause of disease and effective treatment methods that need to be further explored, and achieve the effect of wide application prospects
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[0056] This example provides a DNA fragment derived from Chinese experimental minipigs, which has promoter activity and is denoted as SIECP. For the detection flow chart, see figure 1 .
[0057] 1. Acquisition of target fragments
[0058] Using the Chinese experimental minipig genome as a template, PCR amplification was performed to obtain PCR products. The primers used are as follows:
[0059] The upstream primer is 5'-CGacgcgtGTTTGCTTGGCTTGATCAGCAG-3' (containing MluI recognition sequence);
[0060] The downstream primer is 5'-CTAgctagcATTGACCAGCTCCCCTCTAGTG-3' (containing NheI recognition sequence).
[0061] 2. Construction of pGL3-SIECP recombinant plasmid
[0062] The PCR product obtained in step 1 was double-digested with MluI and NheI, and the resulting fragment was connected to the large vector fragment obtained by double-digestion of the pGL3-Basic plasmid with MluI and NheI to obtain a recombinant plasmid, and the recombinant plasmid with the correct sequence was ...
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