Parallel gene cloning and protein production in multiple expression systems.
To quickly find an optimal expression system for recombinant protein production, a set of vectors with the same restriction sites were constructed for parallel cloning of a target gene and recombinant protein production in prokaryotic and eukaryotic expression systems, simultaneously. These vectors include nucleotide sequences encoding protein tags and protease recognition sites for tag removal, followed by the cloning sites 5'-EcoRI/3'-XhoI identical in these vectors for ligating with the sticky-end PCR product of a target gene. Our vectors allow parallel gene cloning and protein production in multiple expression systems with minimal cloning effort.
Wang
Hui-Min
HM
Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan, ROC.
Shih
Yan-Ping
YP
Hu
Su-Ming
SM
Lo
Wen-Tsung
WT
Lin
Hui-Min
HM
Ding
Shih-Shiu
SS
Liao
Hsin-Chi
HC
Liang
Po-Huang
PH
D016367:Baculoviridae / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D002479:Inclusion Bodies / Q000378:metabolism; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins