Increased expression of transgene in stably transformed cells of Dunaliella salina by matrix attachment regions
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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.
KeywordsChloramphenicol acetyltransferase Dunaliella salina Matrix attachment region Transgene expression
This work was supported by the grants from the National Natural Science Foundation of China (no. 30470030; 30270031) and Special Foundation for Training of Doctoral Students from Institutions of Higher Learning, Ministry of Education of P.R. China (no. 20050459007) and carried out at the Henan Key Laboratory of Molecular Medicine, P.R.C.
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