Abstract
Research studies in the heterologous expression of exogenous genes in living host cells are not new and have been ongoing since the first recombinant DNA molecule was created in the 1970’s [1]. Conventionally, molecular cloning to produce recombinant DNA very often revolve around cleavage of chromosomal DNA or polymerase chain reaction (PCR)-amplified DNA molecules using restriction enzymes and insertion of the DNA fragments of interest with vector DNA, such as plasmids, bacmids or cosmids, with DNA ligase. The recombinant DNA that is constructed is then delivered or transformed into host cells for expression.
Highlights
Molecular cloning to produce recombinant DNA very often revolve around cleavage of chromosomal DNA or polymerase chain reaction (PCR)-amplified DNA molecules using restriction enzymes and insertion of the DNA fragments of interest with vector DNA, such as plasmids, bacmids or cosmids, with DNA ligase
This cloning technology allows the simultaneous insertion of multiple DNA fragments into a single destination vector using site-specific recombinase, the Integrase enzyme, to produce the expression clone [2]
The main reason is that Gateway system utilizes two enzyme mixes, commercially known as BP Clonase and LR Clonase, which are defined by the technology supplier
Summary
Molecular cloning to produce recombinant DNA very often revolve around cleavage of chromosomal DNA or polymerase chain reaction (PCR)-amplified DNA molecules using restriction enzymes and insertion of the DNA fragments of interest with vector DNA, such as plasmids, bacmids or cosmids, with DNA ligase. Many researchers are interested in the co-transformation of multiple DNA elements of interest for expression studies. The idea of incorporating more than one DNA element into a single vector for the ease of transformation is more eminent.
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