Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1984 Jan 25;12(2):1227–1242. doi: 10.1093/nar/12.2.1227

Structure and expression in Escherichia coli of a cloned rat interferon-alpha gene.

R Dijkema, P Pouwels, A de Reus, H Schellekens
PMCID: PMC318569  PMID: 6320120

Abstract

DNA synthesized by in vitro transcription on rat interferon (IFN) mRNA has been cloned and amplified as recombinant DNA. The nucleotide sequence of these rat IFN cDNA clones revealed i. the partial presence of the coding region of the gene and ii all cDNA clones were derived from the same subtype of rat IFN-alpha mRNA. Purified inserted fragments were used as a hybridisation probe against chromosomal "Southern blots" to show that at least twelve rat IFN-alpha-related sequences are present in the genome. A lambda-linked rat gene library was screened with the cDNA probes, resulting in an equivalent number of rat IFN-alpha-related hybrid phages. By use of a 3'-noncoding region as a probe, the chromosomal counterpart of the cDNA clones could be detected and the nucleotide sequence of its coding region has been determined. Expression of the coding region in E. coli yielded biologically active IFN, when tested for in vitro or in vivo antiviral activity.

Full text

PDF
1227

Images in this article

1233Image
on p.1233

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  2. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. Cantell K., Hirvonen S., Mogensen K. E., Pyhälä L. Human leukocyte interferon: production, purification, stability, and animal experiments. In Vitro Monogr. 1974;(3):35–38. [PubMed] [Google Scholar]
  4. Derynck R., Content J., DeClercq E., Volckaert G., Tavernier J., Devos R., Fiers W. Isolation and structure of a human fibroblast interferon gene. Nature. 1980 Jun 19;285(5766):542–547. doi: 10.1038/285542a0. [DOI] [PubMed] [Google Scholar]
  5. Derynck R., Remaut E., Saman E., Stanssens P., De Clercq E., Content J., Fiers W. Expression of human fibroblast interferon gene in Escherichia coli. Nature. 1980 Sep 18;287(5779):193–197. doi: 10.1038/287193a0. [DOI] [PubMed] [Google Scholar]
  6. Gannon F., O'Hare K., Perrin F., LePennec J. P., Benoist C., Cochet M., Breathnach R., Royal A., Garapin A., Cami B. Organisation and sequences at the 5' end of a cloned complete ovalbumin gene. Nature. 1979 Mar 29;278(5703):428–434. doi: 10.1038/278428a0. [DOI] [PubMed] [Google Scholar]
  7. Goeddel D. V., Leung D. W., Dull T. J., Gross M., Lawn R. M., McCandliss R., Seeburg P. H., Ullrich A., Yelverton E., Gray P. W. The structure of eight distinct cloned human leukocyte interferon cDNAs. Nature. 1981 Mar 5;290(5801):20–26. doi: 10.1038/290020a0. [DOI] [PubMed] [Google Scholar]
  8. Goeddel D. V., Yelverton E., Ullrich A., Heyneker H. L., Miozzari G., Holmes W., Seeburg P. H., Dull T., May L., Stebbing N. Human leukocyte interferon produced by E. coli is biologically active. Nature. 1980 Oct 2;287(5781):411–416. doi: 10.1038/287411a0. [DOI] [PubMed] [Google Scholar]
  9. Gray P. W., Goeddel D. V. Structure of the human immune interferon gene. Nature. 1982 Aug 26;298(5877):859–863. doi: 10.1038/298859a0. [DOI] [PubMed] [Google Scholar]
  10. Gray P. W., Leung D. W., Pennica D., Yelverton E., Najarian R., Simonsen C. C., Derynck R., Sherwood P. J., Wallace D. M., Berger S. L. Expression of human immune interferon cDNA in E. coli and monkey cells. Nature. 1982 Feb 11;295(5849):503–508. doi: 10.1038/295503a0. [DOI] [PubMed] [Google Scholar]
  11. Gurdon J. B., Lane C. D., Woodland H. R., Marbaix G. Use of frog eggs and oocytes for the study of messenger RNA and its translation in living cells. Nature. 1971 Sep 17;233(5316):177–182. doi: 10.1038/233177a0. [DOI] [PubMed] [Google Scholar]
  12. Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
  13. Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
  14. Houghton M., Jackson I. J., Porter A. G., Doel S. M., Catlin G. H., Barber C., Carey N. H. The absence of introns within a human fibroblast interferon gene. Nucleic Acids Res. 1981 Jan 24;9(2):247–266. doi: 10.1093/nar/9.2.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. ISAACS A., LINDENMANN J. Virus interference. I. The interferon. Proc R Soc Lond B Biol Sci. 1957 Sep 12;147(927):258–267. doi: 10.1098/rspb.1957.0048. [DOI] [PubMed] [Google Scholar]
  16. Lawn R. M., Adelman J., Franke A. E., Houck C. M., Gross M., Najarian R., Goeddel D. V. Human fibroblast interferon gene lacks introns. Nucleic Acids Res. 1981 Mar 11;9(5):1045–1052. doi: 10.1093/nar/9.5.1045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nagata S., Mantei N., Weissmann C. The structure of one of the eight or more distinct chromosomal genes for human interferon-alpha. Nature. 1980 Oct 2;287(5781):401–408. doi: 10.1038/287401a0. [DOI] [PubMed] [Google Scholar]
  19. Nagata S., Taira H., Hall A., Johnsrud L., Streuli M., Ecsödi J., Boll W., Cantell K., Weissmann C. Synthesis in E. coli of a polypeptide with human leukocyte interferon activity. Nature. 1980 Mar 27;284(5754):316–320. doi: 10.1038/284316a0. [DOI] [PubMed] [Google Scholar]
  20. Ohno S., Taniguchi T. Structure of a chromosomal gene for human interferon beta. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5305–5309. doi: 10.1073/pnas.78.9.5305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sagar A. D., Sehgal P. B., Slate D. L., Ruddle F. H. Multiple human beta interferon genes. J Exp Med. 1982 Sep 1;156(3):744–755. doi: 10.1084/jem.156.3.744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sargent T. D., Wu J. R., Sala-Trepat J. M., Wallace R. B., Reyes A. A., Bonner J. The rat serum albumin gene: analysis of cloned sequences. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3256–3260. doi: 10.1073/pnas.76.7.3256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schellekens H., De Wilde G. A., Weimar W. Production and initial characterization of rat interferon. J Gen Virol. 1980 Jan;46(1):243–247. doi: 10.1099/0022-1317-46-1-243. [DOI] [PubMed] [Google Scholar]
  24. Schellekens H., de Reus A., Bolhuis R., Fountoulakis M., Schein C., Ecsödi J., Nagata S., Weissmann C. Comparative antiviral efficiency of leukocyte and bacterially produced human alpha-interferon in rhesus monkeys. Nature. 1981 Aug 20;292(5825):775–776. doi: 10.1038/292775a0. [DOI] [PubMed] [Google Scholar]
  25. Schellekens H., van Eerd P. M., de Reus A., Weck P. K., Stebbing N. Antiviral and side effects of interferons produced by recombinant DNA techniques as tested in rhesus monkeys. Antiviral Res. 1982 Oct;2(5):313–318. doi: 10.1016/0166-3542(82)90054-7. [DOI] [PubMed] [Google Scholar]
  26. Shaw G. D., Boll W., Taira H., Mantei N., Lengyel P., Weissmann C. Structure and expression of cloned murine IFN-alpha genes. Nucleic Acids Res. 1983 Feb 11;11(3):555–573. doi: 10.1093/nar/11.3.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  28. Taira H., Broeze R. J., Jayaram B. M., Lengyel P., Hunkapiller M. W., Hood L. E. Mouse interferons: amino terminal amino acid sequences of various species. Science. 1980 Feb 1;207(4430):528–530. doi: 10.1126/science.7352261. [DOI] [PubMed] [Google Scholar]
  29. Wieringa B., Mulder J., van der Ende A., Bruggeman A., Ab G., Gruber M. Purification of vitellogenin mRNA and serum albumin mRNA from avian liver by preparative gel electrophoresis. Eur J Biochem. 1978 Aug 15;89(1):67–79. doi: 10.1111/j.1432-1033.1978.tb20897.x. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES