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The use of chimeric gene constructs to express a bacterial endoglucanase in mammalian cells

, : The use of chimeric gene constructs to express a bacterial endoglucanase in mammalian cells. Biochimica et Biophysica Acta 1130(3): 259-266

The synthesis and secrtion of a truncated Clostridium thermocellum endoglucanase (EGE') encoded by the celE' gene was investigated in Chinese hamster ovary (CHO) cells. Fusion genes consisting of the human growth hormone (hGH) gene and celE', transcribed from the SV40 early enhancer/promoter, were constructed and stably transfected into CHO cells. A gene consisting of celE' inserted into the first exon of the hGH gene resulted in the synthesis of truncated proteins (.gtoreq. 22 kDa) lacking endoglucanase activity. Cloning celE' into the second exon of the hGH gene, resulted in the synthesis and secretion of a 50 kDa protein with endoglucanase activity. A 50 kDa protein was also synthesised by cells transfected with celE' cloned into the fifth exon of the hGH gene. However, despite a 5-fold increase in enzyme activity compared to the exon 2 transfected cell line less than 40% of the protein was secreted. Constructs devoid of introns, in which celE' was fused to the SV40 early promoter and to the rabbit .beta.-globin polyadenylation sequence resulted in a 2-18-fold increase in endoglucanase activity compared to the construct containing introns. In addition more than 75% of the synthesised protein was secreted. Analysis of EGE' encoded mRNA from the transfected cell lines suggests that the presence of introns results in the aberrant splicing of message by the use of cryptic splice sites in the celE' gene. These results demonstrate that introns are not required for the efficient expression of a bacterial endoglucanase in mammalian cells, rather introns appear to reduce expression of the encoded protein.

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Accession: 002530216

PMID: 1314095

DOI: 10.1016/0167-4781(92)90438-6

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Teculescu, D.B., 1985: A substitute for "small airway disease"?. Thorax 40(6): 479-479

Jee Jonathan L.; Black Gerald O., 1994: Deposition and diagenesis of Paleogene carbonate rocks, Sarasota Arch and vicinity, northeastern Gulf of Mexico. Abstracts with Programs - Geological Society of America 26(7): 497

Segura, J.José.; Elbourne, A.; Wanless, E.J.; Warr, G.G.; Voïtchovsky, K.; Atkin, R., 2013: Adsorbed and near surface structure of ionic liquids at a solid interface. The structure of solid-ionic liquid (IL) interfaces has been characterised with unprecedented clarity by employing a range of atomic force microscopy (AFM) imaging techniques and tip pressures appropriate for the system under study. Soft contact a...

Suzuki, Y.; Narisawa, K., 1997: Purification and properties of bovine and human holocarboxylase synthetases. Methods in Enzymology 279: 386-393

Paik, C.-Hoon; Kim, S.-Soo, 1996: Relative toxicity of fenpyroximate to the predatory mite, Amblyseius womersleyi (Acarina: Phytoseiidae) and the twospotted spider mite, Tetranychus urticae (Acarina: Tetranychidae). The selectivity index of fenpyroximate to the predatory mite Amblyseius womersleyi and the twospotted spider mite Tetranychus urticae was evaluated. Adult females and eggs of both species were placed on bean leaf discs dipped in several concentrat...

DAVIS, E., 1946: Hereditary clubbing of digits in two families. British Medical Journal 1: 128-128

del CAMPO, J.C., 2014: Secondary local echinococcosis in a residual cavity of an operated pulmonary hydatid cyst. Boletin. Sociedad de Cirugia del Uruguay 21(3): 265-270

PATANE.L.; D.L.CA.V., 1956: Effect of roentgen rays on the development of oostegite in Porcellio laevis Latreille (Crust. Isopoda). Bollettino Della Societa Italiana di Biologia Sperimentale 32(12): 1512-1515

Lipovetskiĭ, B.M., 1986: Physical exercise test on a treadmill as a criterion of the severity of disorders of cardiac rhythm. Klinicheskaia Meditsina 64(7): 139-142

Larsen J.; Engell T., 1977: Levels of estradiol 17 beta in serum during the 1st half of pregnancy and in threatened abortion. Acta Endocrinologica Supplementum 212: 126