Increased expression of transgene in stably transformed cells of Dunaliella salina by matrix attachment regions.
Nuclear matrix attachment regions (MARs) are known to bind specifically to the nuclear scaffold and are thought to influence expression of the transgenes. In our previous studies, a new deoxyribonucleic acid fragment isolated from Dunaliella salina could bind to the nuclear matrix in vitro and had the typical characteristics of MARs. In this study, to investigate effects of MARs on expression of transgenes in the stably transformed cells of D. salina, expression vectors with and without MARs, which contained chloramphenicol acetyltransferase (CAT) reporter gene driven by D. salina ribulose 1,5-bisphosphate carboxylase/oxygenase promoter, were constructed and delivered, respectively, into cells of D. salina by electroporation. Twenty stably transformed colonies of D. salina were randomly picked out, and CAT gene expression was assayed. The results showed that the CAT enzyme of the colonies of D. salina transformed with the expression vector containing MARs averaged out about 4.5-fold higher than those without MARs, while the transgene expression variation among individuals of transformants decreased threefold. The CAT enzyme in the stably transformed lines was not significantly proportional to the gene copy numbers, suggesting that the effects of MARs on transgene expression may not be through increasing the transgene copy numbers.
Wang
Tianyun
T
Laboratory for Cell Biology, The First Affiliated Hospital, Zhengzhou University, 40 Daxue Road, Zhengzhou, Henan, 450052, People's Republic China.
Xue
Lexun
L
Hou
Weihong
W
Yang
Baosheng
B
Chai
Yurong
Y
Ji
Xiang
X
Wang
Yafeng
Y
D001709:Biotechnology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D000460:Chlorophyta / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D018628:Gene Dosage; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D045170:Matrix Attachment Regions; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D014170:Transformation, Genetic
D017931:DNA Primers; D011994:Recombinant Proteins; D015500:Chloramphenicol O-Acetyltransferase