Different vectors used to transform and clone of nonstructural NS1 gene of Influenza B in Escherichia coli
Other Title(s)
استخدام ناقلات مختلفة لنقل و استنساخ الجين غير التركيبي NS1 لفيروس الإنفلونزا ب في بكتريا الايشريكية القولونية
Author
Source
Iraqi Journal of Veterinary Sciences
Issue
Vol. 33, Issue 2 (31 Dec. 2019), pp.329-333, 5 p.
Publisher
University of Mosul College of Veterinary Medicine
Publication Date
2019-12-31
Country of Publication
Iraq
No. of Pages
5
Main Subjects
Topics
Abstract EN
Flu is a highly contagious and common illness caused by influenza A, B, and C viruses.
The aim of the present study was to investigate the transformation and cloning of NS1B gene with pET-32a, pET-32b and pQE-81L in Escherichia coli BL21(DE3) and DH5α.
pUC57-NS1B synthetic gene was transform and clone in Escherichia coli BL21(DE3).
Isolation, single digestion and ligation with pET-32b using HindIII restriction enzyme.
Amplification of recombinant DNA was done with conventional PCR after transformation.
Screening with IPTG of colonies.
Gel electrophoresis was done for each step of cloning after isolation.
Isolation, double digestion and ligation with pET-32a and pQE-81L using SacI, PstI and HindIII respectively.
Recombinant DNA was attempted to be transformed into E.
coli strains BL21 (DE3) and DH5α.
pUC57 plasmid carrying NS1B gene was successful transformed and isolated from E.
coli BL21 (DE3).
Designed primers used for PCR of NS1B showed successful amplification.
First screening of pET-32b-NS1B colonies using white/blue method, cloning NS1B into pET-32b using single restriction digestion with HindIII, pET-32a using double restriction digestion with SacI and HindIII and pQE-81L using double restriction digestion with PstI and HindIII gave unexpected result.
This result may relate to re-ligation of digested vector for single digestion and uncompleted digestion for vectors of double restriction digestion.
Current study has suggested that recombinant NS1B gene can be cloned using single digestion with other expression vectors.
American Psychological Association (APA)
Dawud, Ali Adil. 2019. Different vectors used to transform and clone of nonstructural NS1 gene of Influenza B in Escherichia coli. Iraqi Journal of Veterinary Sciences،Vol. 33, no. 2, pp.329-333.
https://search.emarefa.net/detail/BIM-917142
Modern Language Association (MLA)
Dawud, Ali Adil. Different vectors used to transform and clone of nonstructural NS1 gene of Influenza B in Escherichia coli. Iraqi Journal of Veterinary Sciences Vol. 33, no. 2 (2019), pp.329-333.
https://search.emarefa.net/detail/BIM-917142
American Medical Association (AMA)
Dawud, Ali Adil. Different vectors used to transform and clone of nonstructural NS1 gene of Influenza B in Escherichia coli. Iraqi Journal of Veterinary Sciences. 2019. Vol. 33, no. 2, pp.329-333.
https://search.emarefa.net/detail/BIM-917142
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references : p. 333
Record ID
BIM-917142