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A strategy for in-house production of a positive selection cloning vector from the commercial pJET1.2/blunt cloning vector at minimal cost.


ABSTRACT:

Key message

In-house production of a positive selection cloning vector could be simple, efficient and low cost.

Abstract

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%.

Supplementary information

The online version contains supplementary material available at 10.1007/s13205-022-03289-x.

SUBMITTER: Nawawi O 

PROVIDER: S-EPMC9363543 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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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 O   Abdullah Mohd Puad MP   Yusuf Chong Yu Lok CYL  

3 Biotech 20220809 9


<h4>Key message</h4>In-house production of a positive selection cloning vector could be simple, efficient and low cost.<h4>Abstract</h4>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  ...[more]

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