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Development of a new pCAMBIA binary vector using annealed oligo cloning technique

Research paper by Amir Bidarigh fard, Fatemeh Dehghan Nayeri, Mahdi Habibi Anbouhi

Indexed on: 14 Mar '16Published on: 26 Feb '16Published in: Gene Reports



Abstract

Here a modified version of pCAMBIA1305.1 plasmid with a new multiple cloning site and signal sequence of KDEL for protein secretion was constructed. For this purpose overlapping oligo method was used to obtain a plasmid with new sites downstream of 35S promoter and a KDEL signal sequence. To do this the GUS reporter gene was removed and replaced by a linker fragment containing recognition sites for the restriction enzymes NcoI, BglII, MluI and BstEII. In addition, etanercept gene was introduced into the linker fragment of the modified pCAMBIA1305.1 vector. DNA inserted into this modified vector can be translated in all three reading frames including signal sequence or starting immediately beyond it.

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