Insecticidal proteins and methods for their use

Patent metadata

Field	Value
Publication number	US-10294490-B2
Application number	US-201615274758-A
Country	US
Kind code	B2
Filing date	Sep 23, 2016
Priority date	Jul 26, 2012
Publication date	May 21, 2019
Grant date	May 21, 2019

Abstract

Compositions and methods for controlling pests are provided. The methods involve transforming organisms with a nucleic acid sequence encoding an insecticidal protein. In particular, the nucleic acid sequences are useful for preparing plants and microorganisms that possess insecticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions are insecticidal nucleic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors for subsequent transformation into organisms of interest including plants, as probes for the isolation of other homologous (or partially homologous) genes. The pesticidal proteins find use in controlling, inhibiting growth or killing Lepidopteran, Coleopteran, Dipteran, fungal, Hemipteran and nematode pest populations and for producing compositions with insecticidal activity.

First claim

That which is claimed:

1. A recombinant AflP-1A polypeptide having at least 85% identity to the amino acid sequence of SEQ ID NO: 2, and comprising the amino acid motifs as represented by positions 15-26 of SEQ ID NO: 257, positions 33-53 of SEQ. ID NO: 257, positions 71-84 of SEQ ID NO: 257, and positions 100-107 of SEQ ID NO: 257.

2. The recombinant AfIP-1A polypeptide of claim 1 , wherein the AfIP-1A polypeptide comprises an amino acid sequence of the formula

(SEQ ID NO: 257)

1               5                   10                  15

Met Thr Ala Lys Asp Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala

                20                  25                  30

Xaa Xaa Gln Xaa Xaa Glu Ile Xaa Xaa Phe Xaa Xaa Asn Xaa Xaa

                35                  40                  45

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

                50                  55                  60

Xaa Xaa Xaa Lys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa

                65                  70                  75

Xaa Xaa Ile Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ile Ala Xaa Xaa Xaa

                80                  85                  90

Xaa Xaa Xaa Xaa Ser Ser Xaa Thr Glu Gly Xaa Phe Xaa Xaa Xaa

                95                  100                 105

Xaa Xaa Asp Lys Xaa Xaa Xaa Xaa Tyr Xaa Xaa Asp Cys Xaa Xaa

                110                 115                 120

Xaa Gly Ser Asn Ser Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

                125                 130                135

Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

                140                 145

Xaa Xaa Xaa Xaa Xaa Thr Xaa Xaa Lys Xaa Xaa,

wherein
Xaa at position 6 is Ile, Thr, Leu, Met, Val or Ser;
Xaa at position 7 is Ala, Met, Val, Leu, Ile or Gly;
Xaa at position 8 is Thr, Asp, Ser or Glu;
Xaa at position 9 is Glu, Leu, Gly, Asp, Ala, Ile, Val or Met;
Xaa at position 10 is Glu, Asn, Asp or Gln;
Xaa at position 11 is Ser, Val, Thr, Ile, Leu or Met;
Xaa at position 12 is Lys, Glu, Arg or Asp;
Xaa at position 13 is Ile, Val, Leu or Met;
Xaa at position 14 is Arg, Gln, Lys or Asn;
Xaa at position 16 is Tyr, Gln, Trp, Phe or Asn;
Xaa at position 17 is Ala, Ser, Gly or Thr;
Xaa at position 19 is Trp, Glu, Phe, Ile, His, Asn or Tyr;
Xaa at position 20 is Ile, Val, Ala, Cys, Glu, Phe, Gly, Met, Asn, Gln, Arg, Ser or Thr;
Xaa at position 23 is Thr, Glu, Ala, Ser, Asp or Gly;
Xaa at position 24 is Ile, Leu, Val or Met;
Xaa at position 26 is Val, Ser, Ile, Leu, Met or Thr;
Xaa at position 27 is Val, Glu, Ile, Leu, Met or Asp;
Xaa at position 29 is Ser, Met, Thr, Ile, Leu or Val;
Xaa at position 30 is Asn, Asp, Ser, Glu, Gln or Thr;
Xaa at position 31 is Phe, Ile, Leu, Val or Met;
Xaa at position 32 is Lys, Glu, Arg or Asp;
Xaa at position 33 is Val, Ile, Leu or Met;
Xaa at position 34 is Glu, Lys, Asp or Arg;
Xaa at position 35 is Gly or Asn;
Xaa at position 36 is Ala, Gly, Asp, Glu, Phe, Gly, Ile, Leu, Pro, Gln, Arg, Ser, Thr, Val, Trp or Tyr;
Xaa at position 37 is Tyr, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Pro, Arg, Ser, Thr, Val or Trp;
Xaa at position 38 is Leu, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Gln, Arg, Ser, Thr, Val, Trp or Tyr;
Xaa at position 39 is Arg, Lys, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Asn, Pro, Ser, Thr, Val, Trp or Tyr;
Xaa at position 40 is Trp, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val or Tyr;
Xaa at position 41 is Gly, Cys or Gln;
Xaa at position 42 is Lys, Cys, Glu, His, Leu, Met, Asn, Gln, Arg or Thr;
Xaa at position 43 is Phe, Tyr, Ala, Cys, Glu, Ile, Leu, Met, Gln, Ser, Val or Trp;
Xaa at position 44 is His, Ala, Asp, Glu, Gly, Lys, Leu, Met, Asn, Pro, Glu, Arg, Ser, Thr, Val, Trp;
Xaa at position 45 is Val, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Ser, Thr or Trp;
Xaa at position 46 is Pro, Ala, Cys, Asp, Glu, Gly, His, Lys, Leu, Met, Gln, Arg, Ser, Thr, Val, Trp or Tyr;
Xaa at position 47 is Gly, Leu or Phe;
Xaa at position 48 is Asp, Asn, Glu, Gln, Leu or Phe;
Xaa at position 50 is Asp, Ser, Glu, Thr, Leu or Phe;
Xaa at position 51 is Lys, Asn, Arg, Gln, Leu or Phe;
Xaa at position 52 is Glu, Leu or Phe;
Xaa at position 53 is Ile, Leu or Phe;
Xaa at position 54 is Ser, Thr, Leu or Phe;
Xaa at position 55 is Pro, Ser, Thr, Leu or Phe;
Xaa at position 56 is Ser, Asp, Thr, Glu, Leu;
Xaa at position 57 is Gln, Thr, Glu, Asn, Ser, Asp, Leu or Phe;
Xaa at position 58 is Ile, Val, Leu, Met or Phe;
Xaa at position 60 is Gly, Lys, Ala or Arg;
Xaa at position 61 is Thr, Ile or Phe;
Xaa at position 62 is Ile, Lys, Val, Leu, Met, Arg or Phe;
Xaa at position 64 is Lys, Ser, Glu, Arg, Thr, Asp, Leu or Phe;
Xaa at position 65 is Asp, Ser, Glu, Thr, Leu or Phe;
Xaa at position 66 is Glu, Gly, Asp or Ala;
Xaa at position 67 is Asp, Thr, Glu or Ser;
Xaa at position 68 is Ser, Lys, Thr or Arg;
Xaa at position 69 is Tyr, Ser, Trp, Phe or Thr;
Xaa at position 70 is Thr, Lys, Ser or Arg;
Xaa at position 73 is Ser, Ala, Thr or Gly;
Xaa at position 74 is Cys, Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Ser, Thr or Tyr;
Xaa at position 76 is Arg, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Ser, Thr, Val, Trp or Tyr;
Xaa at position 77 is Glu, Ala, Asp or Gly;
Xaa at position 78 is Asn, Asp, Gln or Glu;
Xaa at position 79 is Ala, Thr, Gly or Ser;
Xaa at position 82 is Gly, Glu, Asn, Trp or Tyr;
Xaa at position 86 is Gly, Lys, Ala or Arg

Assignees

• Pioneer Hi Bred Int
• Du Pont
• E I Du Pont Nemours And Company

Inventors

• Altier Daniel J
• Barry Jennifer K
• Hendrick Carol A
• Liu Lu
• Patten Phillip A
• Perez-Ortega Claudia D
• Schepers Eric J
• Xie Weiping
• Yalpani Nasser
• Zhao Jianzhou
• Zhong Xiaohong
• Zhu Genhai

Classifications

• C07K14/195

  from bacteria · CPC title

• C12N15/8286 Primary

  for insect resistance · CPC title

• C12N15/8207

  by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers · CPC title

• C07K2319/21

  containing a His-tag · CPC title

• C12N15/8205

  Agrobacterium mediated transformation · CPC title