Contribution of regions 3' and 5' to the hIL-5 gene on the expression of rhIL-5 in CHO-cells.
The effect of the presence of the regions 5' and 3' of the hIL-5 gene on the expression of recombinant hIL-5 in CHO-cells was investigated. The 3.2 kb hIL-5 gene-fragment was cloned and four different dexamethasone-inducible rhIL-5 mammalian expression-vectors were constructed, each containing a different part of the gene-fragment. Results indicated that deletion of the region 5' to the gene increases production three-fold whereas a four-fold decrease in production was observed with deletion of the region 3' to the gene. Deletion of both regions increased rhIL-5 production by only one and a half-fold. These results indicate that there are elements in a 928 bp region 3' to the gene, including the 3'-NTR, that have an enhancing or stabilizing effect on expression of hIL-5 in CHO-cells.
van der Westhuizen
F H
FH
Department of Biochemistry, Potchefstroom University for Christian Higher Education, South Africa.bchfhvdw@puknet.puk.ac.za
Pretorius
P J
PJ
de Wet
W J
WJ
D000818:Animals; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D015870:Gene Expression; D006801:Humans; D015848:Interleukin-5 / Q000096:biosynthesis* / Q000235:genetics*; D008322:Mammals; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion; D014162:Transfection
D015848:Interleukin-5; D011994:Recombinant Proteins