Expression of Human Beta-interferon in Namalwa KJM-1 Which Was Adapted to Serum-free Medium

Overview

Journal Cytotechnology

Specialties Biotechnology
Genetics

Date 1990 Mar 1

PMID 1366592

Citations 10

Authors

H Miyaji

T Mizukami

S Hosoi

S Sato

N Fujiyoshi

S Itoh

Affiliations

Soon will be listed here.

Abstract

A Namalwa cell line, KJM-1, which was adapted to serum-free medium is thought to be a very useful host cell line for recombinant DNA technology. Thus, the utility of Namalwa KJM-1 for expression of foreign genes was examined. As a model system human beta-interferon (beta-IFN) gene was engineered for expression in this cell line. For construction of the beta-IFN expression vector pSE1 beta 1-4, the expression vector pAGE107 was constructed and used. It contains simian virus 40 (SV40) early promoter, the rabbit beta-globin RNA processing signals for splicing and polyadenylation, and SV40 early RNA processing signal for polyadenylation. In addition to the above transcription unit, pAGE107 contains the ampicillin-resistance gene and G418-resistance gene. They can confer ampicillin resistance to Escherichia coli (E. coli) and G418 resistance to animal cells. To introduce plasmid DNA into cells, electroporation is a useful method (Wong, 1982; Potter, 1984). We have established conditions for DNA-mediated transfection of Namalwa KJM-1 cell line by electroporation. Among pSE1 beta 1-4-introduced cells, clone 1-3 was further examined for the expression of beta-IFN in serum-free medium. The production level of beta-IFN was elevated with the increase of the cell density. The results indicated that the Namalwa KJM-1 cell line is useful for production of foreign gene products.

Citing Articles

Optimization of cell culture conditions for production of biologically active proteins.

Hosoi S, Miyaji H, Satoh M, Kurimoto T, Mihara A, Fujiyoshi N Cytotechnology. 2012; 5(Suppl 2):17-34.

PMID: 22359044 DOI: 10.1007/BF00573878.

Adaptation of mammalian cells to growth in serum-free media.

Sinacore M, Drapeau D, Adamson S Mol Biotechnol. 2000; 15(3):249-57.

PMID: 10986701 DOI: 10.1385/MB:15:3:249.

Modulation of oligosaccharide structure of a pro-urokinase derivative (pro-UK delta GS1) by changing culture conditions of a lymphoblastoid cell line Namalwa KJM-1 adapted to serum-free medium.

Hosoi S, Satoh M, Higo K, Sugimoto S, Miyaji H, Karasawa A Cytotechnology. 1995; 19(2):125-35.

PMID: 8987501 DOI: 10.1007/BF00749767.

Stabile production of a thrombin resistant pro-urokinase derivative (PRO-UKS1) by Namalwa KJM-1 cells adapted to serum-free medium.

Hosoi S, Satoh M, Miyaji H, Nishi T, Mizukami T, Hasegawa M Cytotechnology. 1995; 19(1):1-10.

PMID: 8987499 DOI: 10.1007/BF00749750.

Efficient expression of pro-urokinase by human lymphoblastoid Namalwa KJM-1 cells using moloney retroviral promoter.

Satoh M, Miyaji H, Nishi T, Mizukami T, Sato S, Itoh S Cytotechnology. 1995; 18(3):167-72.

PMID: 8920107 DOI: 10.1007/BF00767764.

References

OHare K, Benoist C, Breathnach R . Transformation of mouse fibroblasts to methotrexate resistance by a recombinant plasmid expressing a prokaryotic dihydrofolate reductase. Proc Natl Acad Sci U S A. 1981; 78(3):1527-31. PMC: 319164. DOI: 10.1073/pnas.78.3.1527. View

Potter H, Weir L, Leder P . Enhancer-dependent expression of human kappa immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc Natl Acad Sci U S A. 1984; 81(22):7161-5. PMC: 392097. DOI: 10.1073/pnas.81.22.7161. View

Weidle U, Buckel P . Establishment of stable mouse myeloma cells constitutively secreting human tissue-type plasminogen activator. Gene. 1987; 57(1):131-41. DOI: 10.1016/0378-1119(87)90184-3. View

Gralnick H, Williams S, Rick M . Role of carbohydrate in multimeric structure of factor VIII/von Willebrand factor protein. Proc Natl Acad Sci U S A. 1983; 80(9):2771-4. PMC: 393910. DOI: 10.1073/pnas.80.9.2771. View

Toneguzzo F, Hayday A, Keating A . Electric field-mediated DNA transfer: transient and stable gene expression in human and mouse lymphoid cells. Mol Cell Biol. 1986; 6(2):703-6. PMC: 367561. DOI: 10.1128/mcb.6.2.703-706.1986. View

Mizukami T, Itoh S . A new SV40-based vector developed for cDNA expression in animal cells. J Biochem. 1987; 101(5):1307-10. DOI: 10.1093/oxfordjournals.jbchem.a121995. View

Ohno S, Taniguchi T . Structure of a chromosomal gene for human interferon beta. Proc Natl Acad Sci U S A. 1981; 78(9):5305-9. PMC: 348733. DOI: 10.1073/pnas.78.9.5305. View

Kaufman R, Wasley L, Spiliotes A, Gossels S, LATT S, Larsen G . Coamplification and coexpression of human tissue-type plasminogen activator and murine dihydrofolate reductase sequences in Chinese hamster ovary cells. Mol Cell Biol. 1985; 5(7):1750-9. PMC: 367294. DOI: 10.1128/mcb.5.7.1750-1759.1985. View

Wong T, Neumann E . Electric field mediated gene transfer. Biochem Biophys Res Commun. 1982; 107(2):584-7. DOI: 10.1016/0006-291x(82)91531-5. View

10.

Dorai H, Moore G . The effect of dihydrofolate reductase-mediated gene amplification on the expression of transfected immunoglobulin genes. J Immunol. 1987; 139(12):4232-41. View

11.

Vieira J, Messing J . Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985; 33(1):103-19. DOI: 10.1016/0378-1119(85)90120-9. View

12.

Armstrong J . Semi-micro, dye-binding assay for rabbit interferon. Appl Microbiol. 1971; 21(4):723-5. PMC: 377263. DOI: 10.1128/am.21.4.723-725.1971. View

13.

Kaufman R, Sharp P . Amplification and expression of sequences cotransfected with a modular dihydrofolate reductase complementary dna gene. J Mol Biol. 1982; 159(4):601-21. DOI: 10.1016/0022-2836(82)90103-6. View

14.

Hosoi S, Mioh H, Anzai C, Sato S, Fujiyoshi N . Establishment of Namalva cell lines which grow continuously in glutamine-free medium. Cytotechnology. 2012; 1(2):151-8. DOI: 10.1007/BF00146816. View