PT  - JOURNAL ARTICLE
AU  - Zhongtian Liu
AU  - Tingting Zhang
AU  - Kun Xu
TI  - Efficient and flexible strategies for gene cloning and vector construction using overlap PCR
AID  - 10.1101/101147
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 101147
4099  - http://biorxiv.org/content/early/2017/01/18/101147.short
4100  - http://biorxiv.org/content/early/2017/01/18/101147.full
AB  - Gene cloning and vector construction are basic technologies in modern molecular biology for gene functional study. Here, we present flexible and efficient strategies for gene cloning and vector construction using overlap PCR. We firstly cloned the open reading frames (ORFs) of the porcine MSTN, chicken OVA and human α-glucosidase genes by overlap PCR-based assembling of their exons, which could be amplified with genomic DNAs as the templates without RNA extraction and RT-PCR reaction. Secondly, we generated additionally three designed functional cassettes by overlap PCR-based assembling of different DNA elements, which facilitated the construction their expression vectors greatly. Moreover, we further developed an interesting overlap-circled PCR method for fast plasmid vector construction without any cutting and ligating procedure. These advanced applications of overlap PCR provide useful alternative tools for gene cloning and vector construction.