Translation of m-RNA

• US 5,489,527 A
• Filed: 04/12/1993
• Issued: 02/06/1996
• Est. Priority Date: 06/04/1986
• Status: Expired due to Term

• Current Assignee: Copernicus Technology Ltd.
• Original Assignee: Diatech Pty Ltd.
• Inventors: Wilson, Thomas M. A.
• Primary Examiner(s): MOSHER, MARY
• Application Number:
  US08/046,358
• Time in Patent Office: 1,030 Days
• Field of Search: 536/24.1, 435/320.1, 435/252.33, 435/69.1, 435/240.1, 435/240.2, 435/240.4, 435/252.3, 435/172.3, 935/33, 935/34, 935/35, 935/44, 935/45, 935/66
• US Class Current: 435/252.3
• CPC Class Codes: C12N 15/11 DNA or RNA fragments; Modified forms thereof DNA or RNA not used in recombinant technology, C07H21/00; Non-coding nucleic acids having a biological activity
• CPC Class Codes: C12N 15/63 Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 2830/00 Vector systems having a special element relevant for transcription
• CPC Class Codes: C12N 2830/002 inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor
• CPC Class Codes: C12N 2830/55 from bacteria
• CPC Class Codes: C12N 2840/105 enhancing translation
• CPC Class Codes: C12N 2840/60 from viruses
• CPC Class Codes: C12N 9/1033 Chloramphenicol O-acetyltransferase (2.3.1.28)
• CPC Class Codes: C12N 9/12 transferring phosphorus containing groups, e.g. kinases (2.7)
• CPC Class Codes: C12N 9/2434 acting on beta-1,4-glucosidic bonds
• CPC Class Codes: C12N 9/6481 Pepsins (3.4.23.1; 3.4.23.2; 3.4.23.3)
• CPC Class Codes: C12Y 302/01031 Beta-glucuronidase (3.2.1.31)

Abstract

It has been found that the 5'"'"' regions of RNA viruses act as enhancers of translation of mRNA. The omega prime (Ω'"'"') sequence of Tobacco Mosaic Virus, derivatives and portions thereof, and 5'"'"' leader sequences of other RNA viruses can be ligated upstream of an appropriate mRNA or used in the form of cDNA in an expression vector. The translational enhancers are effective in many different types of expression system, for example plant cells, bacteria, and animal cells, and also in in vitro translation.

Claims

1. DNA comprising two operatively linked subsequences, comprising:

i) a first subsequence comprised of a sequence complementary to the Ω

'"'"' sequence of tobacco mosaic virus, and

ii) a second subsequence downstream of said first subsequence and comprised of a coding sequence heterologous to said virus;

wherein, in mRNA produced by transcription of said first and second subsequences, mRNA complementary to said first subsequence is capable of enhancing translation of mRNA complementary to said second subsequence.

2. A cultured cell transformed with the DNA according to claim 1.

3. A cell line derived from a cell according to claim 2.

4. RNA comprising two operatively linked subsequences, comprising

i) a first subsequence comprised of the Ω

'"'"' sequence of tobacco mosaic virus, and

ii) a second subsequence downstream of said first subsequence and comprised of a coding sequence heterologous to said virus;

wherein said first subsequence is capable of enhancing translation of said second sequence.

5. A DNA vector comprising:

i) a first subsequence comprised of a DNA sequence complementary to the Ω

'"'"' sequence of tobacco mosaic virus, and

ii) a second subsequence downstream of said first subsequence and comprised of a coding sequence heterologous to said virus;

wherein said first subsequence is capable of enhancing the translation of mRNA corresponding to said coding sequence heterologous to said virus.