Polypeptides enabling sorting of proteins to vacuoles in plants

• US 5,360,726 A
• Filed: 11/12/1991
• Issued: 11/01/1994
• Est. Priority Date: 09/12/1989
• Status: Expired due to Term

• Current Assignee: Board of Trustees
• Original Assignee: The Board of Trustees of Michigan State University (Michigan State University)
• Inventors: Raikhel, Natasha V.
• Primary Examiner(s): Benzion, Gary
• Assistant Examiner(s): MOSHER, MARY
• Application Number:
  US07/791,930
• Time in Patent Office: 1,085 Days
• Field of Search: 530/324, 530/326, 435/69.1, 435/69.7, 435/172.3, 435/317.1, 435/69.8, 935/47, 935/48, 935/49, 935/51
• US Class Current: 800/287
• CPC Class Codes: C07K 14/42 Lectins, e.g. concanavalin, phytohaemagglutinin
• CPC Class Codes: C12N 15/8279 for biotic stress resistance, pathogen resistance, disease resistance
• CPC Class Codes: C12N 15/8286 for insect resistance
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

A polypeptide enabling sorting of proteins to vacuoles in plants, particularly tobacco is described. The polypeptide has the sequence VFAEAIAANSTLVAE. Without this sequence, the protein is not sorted to the vacuoles. The polypeptide is attached to the C-terminal region of the protein and is particularly useful for sorting of lectins to the vacuole which are insecticidal.

Claims

1. A method of targeting a foreign protein to the vacuole of plant cells which comprises fusing in frame a sequence encoding a polypeptide Val Phe Ala Xaa₁ Ala Ile Xaa₂ Xaa₃ Asn Ser Thr Leu Xaa₄ Xaa₅ Glu as set forth in SEQ ID NO. 6,

wherein Xaa₁ is selected from the group consisting of Gly and Glu,

Xaa₂ and Xaa₃ are selected from the group consisting of Ala and Thr,

Xaa₄ is selected from the group consisting of Val and Leu, and

Xaa₅ is selected from the group consisting of Ala and Gln to a 3'"'"' end of a coding sequence of a foreign protein having an N-terminal signal sequence which mediates translocation to the endoplasmic reticulum and expressing the fused sequence in transgenic plant cells.

2. The method of claim 1 wherein the protein is a lectin.

3. A method of enabling targeting a foreign protein to the vacuole of plant cells which comprises fusing in frame a sequence encoding a polypeptide Val Phe Ala Glu Ala Ile Ala Ala Ash Set Thr Leu Val Ala Glu as set forth in SEQ ID NO. 1 to a 3'"'"' end of a coding sequence of a foreign protein having an N-terminal signal sequence which mediates translocation to the endoplasmic reticulum and expressing the fused sequence in transgenic plant cells.

4. The method of claim 3 wherein the protein is a lectin.

5. The method of claim 4 wherein the lectin is barley lectin.

6. The method of claim 4 wherein the lectin is rice lectin.