Cloning of MSI-78 Gene in Escherichia coli DH5? and Identification of Positive Recombinant
- VernacularTitle:抗菌肽MSI-78基因在大肠埃希菌DH5?中的克隆及阳性重组体鉴定
- Author:
Xinhe SHI
;
Yingfang WANG
;
Zhe GENG
- Publication Type:Journal Article
- Keywords:
Antibacterial peptides;
MSI-78;
Cloning
- From:
Chinese Journal of Nosocomiology
2009;0(13):-
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To clone the MSI-78 gene for the purpose of providing evidence for further studies in prokaryotic expression and activities of antimicrobial peptides. METHODS According to the amino acid sequences of MSI-78,the MSI-78 gene was designed favorable for the Escherichia coli codons. After EcoRⅠand PstⅠ disgestion,cohesive ends were added to both ends respectively and the MSI-78 gene was synthesized by chemical methods. Then,the MSI-78 gene was ligated with pUC-18,transformed into the E. coli DH5?. Through filtration of ? complementary screening,the positive recombinant was finally identified by enzyme digestion of ECORⅠand ECORⅠ/PstⅠ and by PCR. RESULTS The MSI-78 gene was ligated with pUC-18 and transformed into the E. coli DH5?. As a result,MSI-78 gene was cloned in E. coli DH5? successfully. CONCLUSIONS The cloning of the MSI-78 gene provides evidence for further studies of its prokaryotic expression and activities of antimicrobial peptides.
