A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost
DNA cloning technology requires a vector to harbour a gene of interest for multiplication of the gene in bacterial cells. Positive selection vector has become a popular type of cloning vector due to the simplicity and efficiency of the positive selection system. Due to the presence of a toxic gene,...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Published: |
SpringerLink
2022
|
| Online Access: | http://psasir.upm.edu.my/id/eprint/100436/ |
| _version_ | 1848863323427176448 |
|---|---|
| author | Nawawi, Omar Abdullah, Mohd Puad Yusuf, Chong Yu Lok |
| author_facet | Nawawi, Omar Abdullah, Mohd Puad Yusuf, Chong Yu Lok |
| author_sort | Nawawi, Omar |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | DNA cloning technology requires a vector to harbour a gene of interest for multiplication of the gene in bacterial cells. Positive selection vector has become a popular type of cloning vector due to the simplicity and efficiency of the positive selection system. Due to the presence of a toxic gene, propagation of a commercial positive selection vector in common laboratory E. coli strains is infeasible. This study demonstrated a strategy for propagation and in-house production of a commercial positive selection vector, i.e., pJET1.2/blunt cloning vector, at low cost. This was done by insertion of a specially designed DNA fragment (MCS fragment), which can be easily removed later by EcoRV digestion, into the pJET1.2/blunt cloning vector to allow the propagation of the modified plasmid (termed pJET1.2M) in common E. coli strains. Removal of the MCS fragment from the pJET1.2M plasmid produces the pJET1.2/blunt cloning vector ready for gene cloning. The self-made pJET1.2/blunt cloning vector exhibited a cloning efficiency of 100%. |
| first_indexed | 2025-11-15T13:31:06Z |
| format | Article |
| id | upm-100436 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-15T13:31:06Z |
| publishDate | 2022 |
| publisher | SpringerLink |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-1004362023-12-15T23:18:08Z http://psasir.upm.edu.my/id/eprint/100436/ A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost Nawawi, Omar Abdullah, Mohd Puad Yusuf, Chong Yu Lok DNA cloning technology requires a vector to harbour a gene of interest for multiplication of the gene in bacterial cells. Positive selection vector has become a popular type of cloning vector due to the simplicity and efficiency of the positive selection system. Due to the presence of a toxic gene, propagation of a commercial positive selection vector in common laboratory E. coli strains is infeasible. This study demonstrated a strategy for propagation and in-house production of a commercial positive selection vector, i.e., pJET1.2/blunt cloning vector, at low cost. This was done by insertion of a specially designed DNA fragment (MCS fragment), which can be easily removed later by EcoRV digestion, into the pJET1.2/blunt cloning vector to allow the propagation of the modified plasmid (termed pJET1.2M) in common E. coli strains. Removal of the MCS fragment from the pJET1.2M plasmid produces the pJET1.2/blunt cloning vector ready for gene cloning. The self-made pJET1.2/blunt cloning vector exhibited a cloning efficiency of 100%. SpringerLink 2022-08-09 Article PeerReviewed Nawawi, Omar and Abdullah, Mohd Puad and Yusuf, Chong Yu Lok (2022) A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost. 3 Biotech, 12. art. no. 216. pp. 1-5. ISSN 2190-5738 https://link.springer.com/article/10.1007/s13205-022-03289-x 10.1007/s13205-022-03289-x |
| spellingShingle | Nawawi, Omar Abdullah, Mohd Puad Yusuf, Chong Yu Lok A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost |
| title | A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost |
| title_full | A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost |
| title_fullStr | A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost |
| title_full_unstemmed | A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost |
| title_short | A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost |
| title_sort | strategy for in-house production of a positive selection cloning vector from the commercial pjet1.2/blunt cloning vector at minimal cost |
| url | http://psasir.upm.edu.my/id/eprint/100436/ http://psasir.upm.edu.my/id/eprint/100436/ http://psasir.upm.edu.my/id/eprint/100436/ |