Biopolym. Cell. 2006; 22(3):210-216.
Genome and Its Regulation
Study of transient expression of human apoA-1 gene under control of various promoters in CHO-K1 cells
1Gilchuk Y. M., 1Sukhorada O. M., 1Ruban T. A., 1Toporova O. K., 1Kordyum V. A.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680


The recombinant plasmids containing the genomic human apoA-1 gene under the transcriptional control of hybrid promoters, namely, the cytomegalovirus immediate early enhancer/chicken β-actin promoter and first intron (CAG promoter) and cytomegalovirus immediate early enhancer/promoter with intron A (hCMVintronA promoter) have been constructed. The transient expression of human apoA-1 gene has been studied in CHO-K1 cells using designed and pTRapo vectors. The expression of the transgene for transfected with pTRapo and pTRapochCMVintronA CHO-K1 cells have been shown.
Keywords: transient expression, apolipoprotein A-1, CHO-K1


[1] Romano G, Michell P, Pacilio C, Giordano A. Latest developments in gene transfer technology: achievements, perspectives, and controversies over therapeutic applications. Stem Cells. 2000;18(1):19-39.
[2] Bogdanenko EV, Sviridov IuV, Moskovtsev AA, Zhdanov RI. Nonviral gene transfer in vivo in gene therapy. Vopr Med Khim. 2000;46(3):226-45.
[3] Nishikawa M, Hashida M. Nonviral approaches satisfying various requirements for effective in vivo gene therapy. Biol Pharm Bull. 2002;25(3):275-83.
[4] Wurm FM. Production of recombinant protein therapeutics in cultivated mammalian cells. Nat Biotechnol. 2004;22(11):1393-8.
[5] Xia W, Bringmann P, McClary J, Jones PP, Manzana W, Zhu Y, Wang S, Liu Y, Harvey S, Madlansacay MR, McLean K, Rosser MP, MacRobbie J, Olsen CL, Cobb RR. High levels of protein expression using different mammalian CMV promoters in several cell lines. Protein Expr Purif. 2006;45(1):115-24.
[6] Toporova OK, Novikova SN, Lihacheva LI, Suhorada OM, Ruban TA, Kozel JA, Irodov DM, Kordium VA. Non-viral gene delivery of human apoA1 into mammalian cells in vitro and in vivo. Biopolym Cell. 2004; 20(1-2):25-32.
[7] Kordyum VA, Toporova EK, Okunev OV, Pokholenko YA, Suchorada EM, Ruban TA, Andrienko VI, Irodov DM. Novel cell line with multiple markers, derivative of CHO-K1. Biopolym. Cell. 2003; 19(3):252-6.
[8] Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Lab. press, 1989. Vol. 1.
[9] Ausubel FM, Brent R, Kingston RE, Moore DD, SeidmanJ. G., Smith J. A, StruhlK. Current protocols in molecular biology. New York: John Wiley and Sons, 1997. Vol. 1.
[10] Boussif O, Lezoualc'h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, Behr JP. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci U S A. 1995;92(16):7297-301.
[11] Bollag DM, Rozychi MD. Protein Methods. New York: Wiley-Liss. Inc., 1996. 415 p.
[12] The Protein Protocols Handbook. Ed. J. M. Walker. Totowa, New Jersey: Humana press Inc., 2002. 1139 p.
[13] Boshart M, Weber F, Jahn G, Dorsch-H?sler K, Fleckenstein B, Schaffner W. A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell. 1985;41(2):521-30.
[14] Fitzsimons HL, Bland RJ, During MJ. Promoters and regulatory elements that improve adeno-associated virus transgene expression in the brain. Methods. 2002;28(2):227-36.
[15] Palmiter RD, Sandgren EP, Avarbock MR, Allen DD, Brinster RL. Heterologous introns can enhance expression of transgenes in mice. Proc Natl Acad Sci U S A. 1991;88(2):478-82.
[16] Chapman BS, Thayer RM, Vincent KA, Haigwood NL. Effect of intron A from human cytomegalovirus (Towne) immediate-early gene on heterologous expression in mammalian cells. Nucleic Acids Res. 1991;19(14):3979-86.
[17] Niwa H, Yamamura K, Miyazaki J. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene. 1991;108(2):193-9.
[18] Makrides SC. Components of vectors for gene transfer and expression in mammalian cells. Protein Expr Purif. 1999;17(2):183-202.
[19] Xu ZL, Mizuguchi H, Ishii-Watabe A, Uchida E, Mayumi T, Hayakawa T. Optimization of transcriptional regulatory elements for constructing plasmid vectors. Gene. 2001;272(1-2):149-56.