Bombyx mori posterior silk gland bioreactor dual-promoter universal plasmid for expressing T4 ligase and application and method thereof
A bioreactor, dual-promoter technology, applied in biochemical equipment and methods, ligase, introduction of foreign genetic material using vectors, etc. in reaching etc.
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[0031] The present invention will be further described below in conjunction with drawings and embodiments.
[0032] Embodiments of the present invention are as follows:
[0033] A) preparation of the plasmid of the present invention:
[0034]Use the 5' end sequence of T4 ligase with ApaI restriction site sequence and the 3' end sequence of T4 ligase with NheI restriction site sequence as primers to contain A3 gene promoter-green fluorescent protein gene-SV40 - sericin gene promoter-T4 ligase gene-SV40 ([A3-EGFP-SV40]-[Serpromoter-T4Ligase-SV40]) piggy-10522 plasmid (its base sequence such as SEQ ID NO.2) is used as a template to obtain A T4 ligase gene with a length of 1536bp (its base sequence is as shown in SEQ ID NO.3), and the 5' end of the sequence contains an ApaI restriction site, and the 3' end contains an NheI restriction site.
[0035] Then use ApaI and NheI to double-digest the double-promoter plasmid piggy-8480 plasmid (its base sequence is as SEQ ID NO.4) in the...
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