Construction method of recombinant vector, constructed recombinant vector and application of recombinant vector
A recombinant vector and construction method technology, applied in the fields of molecular biology and protein, can solve the problems of tedious preparation work and double the cost, and achieve the effects of improving transfection efficiency, reducing workload and high-efficiency expression.
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[0116] 1. Construction of IL6 antibody 1A5 sequence eukaryotic expression vector
[0117]The phage clone 1A5 of the anti-hIL-6 protein ScFv antibody with high affinity (the nucleotide sequence of the 1A5 heavy chain is shown in SEQ ID NO: 6, and its amino acid sequence is shown in SEQ ID NO: 7; the core of 1A5-VH The nucleotide sequence is shown in SEQ ID NO: 8, and its amino acid sequence is shown in SEQ ID NO: 9; the nucleotide sequence of the 1A5 light chain is shown in SEQ ID NO: 10, and its amino acid sequence is shown in SEQ ID NO: 11 shown; the nucleotide sequence of 1A5-VL is as shown in SEQ ID NO: 12, and its amino acid sequence is as shown in SEQ ID NO: 13) as template, respectively design primer 1A5-VL upstream primer / 1A5-VL downstream primer amplification The light chain variable region 1A5-VL and the primer 1A5-VH upstream primer / 1A5-VH downstream primer amplify the DNA fragment of the heavy chain variable region 1A5-VH. PCR amplification was performed with prime...
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