|
|
fbook(金币+100, 翻译EPI+1): 2010-09-30 18:04:39
To detect antimicrobial activity,we take amino acid sequence of Bovine antimicrobial peptide (Indolicidin) as a template, adding Kallikrein protease cleavage sites to the C terminal of peptide to get new antimicrobial peptide NK-16.
Based on amino acid sequence and E.coli preferred codons,four overlapping fragments were backward translated ,designed and synthesized.
The restriction sites Xho I and Nco I which were used to express the contructino of vector were introduced onto 5 'and 3'terminal.
Overlapping PCR technique was used to connect synthetic fragment and made the connected nucleotide sequence cloned into pET 30a (+) vector to construct multi-carrier PET30a (+)-NK-16.the multiple expression vector PET30a (+)-NK-16 was proved to be constructed successfully by the identification by Double digestion and PCR. the right construction of multiple expression vector PET30a (+)-NK-16 lay the groundwork for the highl expression of prokaryotic antimicrobial peptides. |
|
回复此楼