Transcription Factors, DNA and Methods for Introduction of Value-Added Seed Traits and Stress Tolerance

Abstract

Abscisic acid-inducible gene expression in different plant tissues is enhanced synergistically by the co-expression of a B3-domain transcription factor and various bZIP-domain transcription factors, or a different B3-domain transcription factor. Using these transcription factors in novel formulations, as shown by examples, will confer value-added traits to transgenic plants, including, but not limited to, higher levels of heterologous gene expression, drought and salt tolerance, viability and productivity under stress, and enhanced nutrient reserves and seed properties.

1 . A method of producing a transgenic plant comprising
 a) transforming a plant cell with an expression vector comprising
 i) a B3-domain transcription factor having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID NO:1 or SEQ. ID. NO.: 3 and 
 ii) a bZIP transcription factor having a polynucleotide sequence with a bZIP domain which is at least 25% identical to the bZIP domain of SEQ. ID NO: 5, and 
   b) generating from the plant cell a transgenic plant with an increased tolerance to environmental stress as compared to a wild type variety of the plant.   
     
     
         2 . The method of  claim 1 , wherein said B3-domain transcription factor is VP1 as defined by SEQ.ID.NO.: 1. 
     
     
         3 . The method of  claim 2 , wherein said bZIP domain transcription factor is ABI5 as defined by SEQ.ID.NO.: 5 or ABF1 as defined by SEQ.ID.NO.: 7. 
     
     
         4 . The method of  claim 1 , wherein said bZIP domain transcription factor is ABI5 as defined by SEQ.ID.NO.: 5 or ABF1 as defined by SEQ.ID.NO.: 7. 
     
     
         5 . The method of  claim 1 , wherein said bZIP domain transcription factor is selected from a group of polynucleotides consisting of SEQ.ID.NO. 9, SEQ.ID.NO. 11, SEQ.ID.NO. 15, SEQ.ID.NO. 17, SEQ.ID.NO.19, SEQ.ID.NO. 21, SEQ.ID.NO. 23, SEQ.ID.NO.25, and SEQ.ID.NO. 27. 
     
     
         6 . The method of  claim 1 , wherein said B3 domain transcription factors are selected from a group of polynucleotides consisting of SEQ.ID.NO. 29, SEQ.ID.NO.31, SEQ.ID.NO. 33, SEQ.ID.NO. 35, SEQ.ID.NO. 37, SEQ.ID.NO. 39, SEQ.ID.NO. 41, SEQ.ID.NO. 43, SEQ.ID.NO.45, SEQ.ID.NO. 47, SEQ.ID.NO. 49, SEQ.ID.NO.51, SEQ.ID.NO. 53, SEQ.ID.NO. 55, SEQ.ID.NO. 57, SEQ.ID.NO. 59, SEQ.ID.NO. 61 SEQ.ID.NO. 63, and SEQ.ID.NO.65. 
     
     
         7 . The method of  claim 1 , wherein said transgenic plant is a monocot plant or a dicot plant. 
     
     
         8 . The method of  claim 1 , wherein said expression vector is a tissue-specific expression vector. 
     
     
         9 . A method of producing a transgenic plant comprising
 a) transforming a plant cell with an expression vector comprising
 i) a B3-domain transcription factor having a polypeptide sequence with a B3 domain which is at least 40% identical to the B3 domain of SEQ. ID NO:2 or SEQ. ID. NO.: 4 and 
 ii) a bZIP transcription factor having a polypeptide sequence with a bZIP domain which is at least 40% identical to the bZIP domain of SEQ. ID NO: 6, and 
   b) generating from the plant cell a transgenic plant with an increased tolerance to environmental stress as compared to a wild type variety of the plant.   
     
     
         10 . The method of  claim 9 , wherein said B3-domain transcription factor is VP1 as defined by SEQ.ID.NO.: 2. 
     
     
         11 . The method of  claim 10 , wherein said bZIP domain transcription factor is ABI5 as defined by SEQ.ID.NO.: 6 or ABF1 as defined by SEQ.ID.NO.: 8. 
     
     
         12 . The method of  claim 9 , wherein said bZIP domain transcription factor is ABI5 as defined by SEQ.ID.NO.: 6 or ABF1 as defined by SEQ.ID.NO.: 8. 
     
     
         13 . The method of  claim 9 , wherein said bZIP domain transcription factor is selected from a group of polypeptides consisting of SEQ.ID.NO. 10, SEQ.ID.NO. 12, SEQ.ID.NO. 16, SEQ.ID.NO. 18, SEQ.ID.NO.20, SEQ.ID.NO. 22, SEQ.ID.NO. 24, SEQ.ID.NO.26, and SEQ.ID.NO. 28. 
     
     
         14 . The method of  claim 9 , wherein said B3 domain transcription factors are selected from a group of polypeptides consisting of SEQ.ID.NO. 30, SEQ.ID.NO.32, SEQ.ID.NO. 34, SEQ.ID.NO. 36, SEQ.ID.NO. 38, SEQ.ID.NO. 40, SEQ.ID.NO. 42, SEQ.ID.NO. 44, SEQ.ID.NO.46, SEQ.ID.NO. 48, SEQ.ID.NO. 50, SEQ.ID.NO.52, SEQ.ID.NO. 54, SEQ.ID.NO. 56, SEQ.ID.NO. 58, SEQ.ID.NO. 60, SEQ.ID.NO. 62 SEQ.ID.NO. 64, and SEQ.ID.NO.66. 
     
     
         15 . The method of  claim 9 , wherein said transgenic plant is a monocot plant or a dicot plant. 
     
     
         16 . The method of  claim 9 , wherein said expression vector is a tissue-specific expression vector. 
     
     
         17 . A method of producing a transgenic plant comprising
 a) transforming a plant cell with an expression vector comprising
 i) a first B3-domain transcription factor having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID NO:1 and 
 ii) a second B3-domain transcription factor having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID NO: 3, and 
   b) generating from the plant cell a transgenic plant with an increased tolerance to environmental stress as compared to a wild type variety of the plant.   
     
     
         18 . The method of  claim 17 , wherein said first B3 domain transcription factor is VP1 as defined by SEQ. ID. NO.: 1. 
     
     
         19 . The method of  claim 18 , wherein said second B3 domain transcription factor is RAV2 as defined by SEQ. ID. NO.: 3. 
     
     
         20 . The method of  claim 17 , wherein said first B3 domain transcription factor is RAV2 as defined by SEQ. ID. NO.: 3. 
     
     
         21 . The method of  claim 17 , wherein said first B3 domain transcription factor or said second B3 domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO.: 29. SEQ. ID. NO.:31, SEQ. ID. NO.:33, SEQ. ID. NO.:35, SEQ. ID. NO.:37, SEQ. ID. NO.:39, SEQ. ID. NO.:41, SEQ. ID. NO.:43, SEQ. ID. NO.:45, SEQ. ID. NO.:47, SEQ. ID. NO.:49, SEQ. ID. NO.:51, SEQ. ID. NO.:53, SEQ. ID. NO.:55, SEQ. ID. NO.:57, SEQ. ID. NO.:59, SEQ. ID. NO.:61, SEQ. ID. NO.:63 and SEQ. ID. NO.:65. 
     
     
         22 . The method of  claim 17 , wherein said transgenic plant is a monocot plant or a dicot plant. 
     
     
         23 . The method of  claim 17 , wherein said expression vector is a tissue-specific expression vector. 
     
     
         24 . A method of producing a transgenic plant comprising
 a) transforming a plant cell with an expression vector comprising
 i) a first B3-domain transcription factor having a polypeptide sequence with a B3 domain which is at least 40% identical to the B3 domain of SEQ. ID NO:2 and 
 ii) a second B3-domain transcription factor having a polypeptide sequence with a B3 domain which is at least 40% identical to the B3 domain of SEQ. ID NO: 4, and 
   b) generating from the plant cell a transgenic plant with an increased tolerance to environmental stress as compared to a wild type variety of the plant.   
     
     
         25 . The method of  claim 24 , wherein said first B3-domain transcription factor is VP1 as defined by SEQ.ID.NO.: 2. 
     
     
         26 . The method of  claim 25 , wherein said second B3-domain transcription factor is RAV2 as defined by SEQ.ID.NO.: 4. 
     
     
         27 . The method of  claim 24 , wherein said first B3 domain transcription factor is RAV2 as defined by SEQ. IS. NO.: 4. 
     
     
         28 . The method of  claims 24 , wherein said first B3 domain transcription factor or said second B3 domain transcription factor is selected from a group of polypeptides consisting of SEQ.ID.NO.: 30, SEQ.ID.NO.:32, SEQ.ID.NO.:34, SEQ.ID.NO.:36, SEQ. ID.NO.:38, SEQ.ID.NO.:40, SEQ.ID.NO.:42, SEQ.ID.NO.:44, SEQ.ID.NO.:46, SEQ. ID.NO.:48, SEQ.ID.NO.:50, SEQ.ID.NO.:52, SEQ.ID.NO.:54, SEQ.ID.NO.:56, SEQ. ID.NO.:58, SEQ.ID.NO.:60, SEQ.ID.NO.:62, SEQ.ID.NO.:64, and SEQ.ID.NO.:66. 
     
     
         29 . The method of  claim 24 , wherein said transgenic plant is a monocot plant or a dicot plant. 
     
     
         30 . The method of  claim 24 , wherein said expression vector is a tissue-specific expression vector. 
     
     
         31 . A transgenic plant, comprising
 a) a B3-domain transcription factor having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID. NO: 1 or SEQ. ID. NO. 3 operably linked to a tissue specific promoter, and   b) a bZIP domain transcription factor having a polynucleotide sequence with a bZIP domain which is at least 25% identical to the bZIP domain of SEQ. ID. NO.:5 operably linked to a tissue specific promoter,   wherein expression of the B3-domain transcription factor and the bZIP domain transcription factor in the tissue of said plant is effective to activate transcription of a gene operably linked to a promoter with which said transcription factor interacts.   
     
     
         32 . The transgenic plant of  claim 31 , wherein said B3-domain transcription factor is VP1 as defined by SEQ.ID.NO.: 1. 
     
     
         33 . The transgenic plant of  claim 32 , wherein said bZIP domain transcription factor is ABI5 as defined by SEQ.ID.NO.: 5 or ABF1 as defined by SEQ.ID.NO.: 7. 
     
     
         34 . The transgenic plant of  claim 31 , wherein said bZIP domain transcription factor is ABI5 as defined by SEQ.ID.NO.: 5 or ABF1 as defined by SEQ.ID.NO.: 7. 
     
     
         35 . The transgenic plant of  claim 31 , wherein said bZIP domain transcription factor is selected from a group of polynucleotides consisting of SEQ.ID.NO. 9, SEQ.ID.NO. 11, SEQ.ID.NO. 15, SEQ.ID.NO. 17, SEQ.ID.NO.19, SEQ.ID.NO. 21, SEQ.ID.NO. 23, SEQ.ID.NO.25, and SEQ.ID.NO. 27. 
     
     
         36 . The transgenic plant of  claim 31 , wherein said B3 domain transcription factors are selected from a group of polynucleotides consisting of SEQ.ID.NO. 29, SEQ.ID.NO.31, SEQ.ID.NO. 33, SEQ.ID.NO. 35, SEQ.ID.NO. 37, SEQ.ID.NO. 39, SEQ.ID.NO. 41, SEQ.ID.NO. 43, SEQ.ID.NO.45, SEQ.ID.NO. 47, SEQ.ID.NO. 49, SEQ.ID.NO.51, SEQ.ID.NO. 53, SEQ.ID.NO. 55, SEQ.ID.NO. 57, SEQ.ID.NO. 59, SEQ.ID.NO. 61 SEQ.ID.NO. 63, and SEQ.ID.NO.65. 
     
     
         37 . The transgenic plant of  claim 31 , wherein said transgenic plant is a monocot plant or a dicot plant. 
     
     
         38 . A transgenic plant, comprising
 a) a B3-domain transcription factor having a polypeptide sequence with a B3 domain which is at least 40% identical to the B3 domain of SEQ. ID. NO:2 or SEQ. ID. NO. 4 operably linked to a tissue specific promoter, and   b) a bZIP domain transcription factor having a polypeptide sequence with a bZIP domain which is at least 40% identical to the bZIP domain of SEQ. ID. NO.:6 operably linked to a tissue specific promoter,   wherein expression of the B3-domain transcription factor and the bZIP domain transcription factor in the tissue of said plant is effective to activate transcription of a gene operably linked to a promoter with which said transcription factor interacts.   
     
     
         39 . The transgenic plant of  claim 38 , wherein said B3-domain transcription factor is VP1 as defined by SEQ.ID.NO.: 2. 
     
     
         40 . The transgenic plant of  claim 39 , wherein said bZIP domain transcription factor is ABI5 as defined by SEQ.ID.NO.: 6 or ABF1 as defined by SEQ.ID.NO.: 8. 
     
     
         41 . The transgenic plant of  claim 38 , wherein said bZIP domain transcription factor is ABI5 as defined by SEQ.ID.NO.: 6 ABF1 as defined by SEQ.ID.NO.: 8. 
     
     
         42 . The transgenic plant of  claim 38 , wherein said bZIP domain transcription factor is selected from a group of polypeptides consisting of SEQ.ID.NO. 10, SEQ.ID.NO. 12, SEQ.ID.NO. 16, SEQ.ID.NO. 18, SEQ.ID.NO.20, SEQ.ID.NO. 22, SEQ.ID.NO. 24, SEQ.ID.NO.26, and SEQ.ID.NO. 28. 
     
     
         43 . The transgenic plant of  claim 38 , wherein said B3 domain transcription factors are selected from a group of polypeptides consisting of SEQ.ID.NO. 30, SEQ.ID.NO.32, SEQ.ID.NO. 34, SEQ.ID.NO. 36, SEQ.ID.NO. 38, SEQ.ID.NO. 40, SEQ.ID.NO. 42, SEQ.ID.NO. 44, SEQ.ID.NO.46, SEQ.ID.NO. 48, SEQ.ID.NO. 50, SEQ.ID.NO.52, SEQ.ID.NO. 54, SEQ.ID.NO. 56, SEQ.ID.NO. 58, SEQ.ID.NO. 60, SEQ.ID.NO. 62 SEQ.ID.NO. 64, and SEQ.ID.NO.66. 
     
     
         44 . The transgenic plant of  claim 38 , wherein said transgenic plant is a monocot plant or a dicot plant. 
     
     
         45 . A transgenic plant, comprising
 a) a first B3-domain transcription factor having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID. NO: 1 operably linked to a tissue specific promoter, and   b) a second B3-domain domain transcription factor having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID. NO.:3 operably linked to a tissue specific promoter,   wherein expression of the first B3-domain transcription factor and the second B3 domain transcription factor in the tissue of said plant is effective to activate transcription of a gene operably linked to a promoter with which said transcription factor interacts.   
     
     
         46 . The transgenic plant of  claim 45 , wherein said first B3 domain transcription factor is VP1 as defined by SEQ. ID. NO.: 1. 
     
     
         47 . The transgenic plant of  claim 46 , wherein said second B3 domain transcription factor is RAV2 as defined by SEQ. ID. NO.: 3. 
     
     
         48 . The transgenic plant of  claim 45 , wherein said first B3 domain transcription factor is RAV2 as defined by SEQ. ID. NO.: 3. 
     
     
         49 . The transgenic plant of  claim 45 , wherein said first B3 domain transcription factor or said second B3 domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO.: 29. SEQ. ID. NO.:31, SEQ. ID. NO.:33, SEQ. ID. NO.:35, SEQ. ID. NO.:37, SEQ. ID. NO.:39, SEQ. ID. NO.:41, SEQ. ID. NO.:43, SEQ. ID. NO.:45, SEQ. ID. NO.:47, SEQ. ID. NO.:49, SEQ. ID. NO.:51, SEQ. ID. NO.:53, SEQ. ID. NO.:55, SEQ. ID. NO.:57, SEQ. ID. NO.:59, SEQ. ID. NO.:61, SEQ. ID. NO.:63 and SEQ. ID. NO.:65. 
     
     
         50 . The transgenic plant of  claim 45 , wherein said transgenic plant is a monocot plant or a dicot plant. 
     
     
         51 . A transgenic plant, comprising
 a) a first B3-domain transcription factor having a polypeptide sequence with a B3 domain which is at least 40% identical to the B3 domain of SEQ. ID. NO:2 operably linked to a tissue specific promoter, and   b) a second B3-domain domain transcription factor having a polypeptide sequence with a B3 domain which is at least 40% identical to the B3 domain of SEQ. ID. NO.:4 operably linked to a tissue specific promoter,   wherein expression of the first B3-domain transcription factor and the second B3 domain transcription factor in the tissue of said plant is effective to activate transcription of a gene operably linked to a promoter with which said transcription factor interacts.   
     
     
         52 . The transgenic plant of  claim 51 , wherein said first B3-domain transcription factor is VP1 as defined by SEQ.ID.NO.: 2. 
     
     
         53 . The transgenic plant of  claim 52 , wherein said second B3-domain transcription factor is RAV2 as defined by SEQ.ID.NO.: 4. 
     
     
         54 . The transgenic plant of  claim 51 , wherein said first B3 domain transcription factor is RAV2 as defined by SEQ. IS. NO.: 4. 
     
     
         55 . The transgenic plant of  claims 51 , wherein said first B3 domain transcription factor or said second B3 domain transcription factor is selected from a group of polypeptides consisting of SEQ.ID.NO.: 30, SEQ.ID.NO.:32, SEQ.ID.NO.:34, SEQ.ID.NO.:36, SEQ. ID.NO.:38, SEQ.ID.NO.:40, SEQ.ID.NO.:42, SEQ.ID.NO.:44, SEQ.ID.NO.:46, SEQ. ID.NO.:48, SEQ.ID.NO.:50, SEQ.ID.NO.:52, SEQ.ID.NO.:54, SEQ.ID.NO.:56, SEQ. ID.NO.:58, SEQ.ID.NO.:60, SEQ.ID.NO.:62, SEQ.ID.NO.:64, and SEQ.ID.NO.:66. 
     
     
         56 . The transgenic plant of  claim 51 , wherein said transgenic plant is a monocot plant or a dicot plant. 
     
     
         57 . A method of screening for a polynucleotide which encodes a B3 domain protein or a bZIP domain protein comprising,
 a) hybridizing a polynucleotide having at least 25% identical to the B3 domain of SEQ. ID. NO:1 or SEQ. ID. NO.: 3 or a polynucleotide having at least 25% identity to the bZIP domain of SEQ. ID. NO. 5: to the polynucleotide to be screened;   b) expressing the polynucleotide to produce a protein; and   c) detecting the presence or absence of B3 domain activity or bZIP domain activity in said protein.   
     
     
         58 . A vector comprising an isolated polynucleotide having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID. NO. 1 or SEQ. ID. NO.: 3 and an isolated polynucleotide having a polynucleotide sequence with a bZIP domain which is at least 25% identical to the bZIP domain of SEQ. ID. NO.: 5. 
     
     
         59 . The vector of  claim 58 , wherein said B3-domain transcription factor is VP1 as defined by SEQ.ID.NO.: 1. 
     
     
         60 . The vector of  claim 59 , wherein said bZIP domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO. 5, and SEQ. ID. NO. 7. 
     
     
         61 . The vector of  claim 58 , wherein said bZIP domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO. 5. and SEQ. ID. NO. 7. 
     
     
         62 . The vector of  claim 58  wherein said bZIP domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO. 9, SEQ. ID. NO. 11, SEQ. ID. NO. 15, SEQ. ID. NO. 17, SEQ. ID. NO. 19, SEQ. ID. NO. 21, SEQ. ID. NO. 23, SEQ. ID. NO. 25 and SEQ. ID. NO. 27. 
     
     
         63 . The vector of  claim 58  wherein said B3 domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO. 29, SEQ. ID. NO. 31, SEQ. ID. NO. 33, SEQ. ID. NO. 35, SEQ. ID. NO. 37, SEQ. ID. NO. 39, SEQ. ID. NO. 41, SEQ. ID. NO. 43, SEQ. ID. NO. 45, SEQ. ID. NO. 47, SEQ. ID. NO. 49, SEQ. ID. NO. 51, SEQ. ID. NO. 53, SEQ. ID. NO. 55, SEQ. ID. NO. 57, SEQ. ID. NO. 59, SEQ. ID. NO. 61, SEQ. ID. NO. 63, and SEQ. ID. NO. 65. 
     
     
         64 . A host cell comprising an isolated polynucleotide having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID. NO. 1 or SEQ. ID. NO.: 3 and an isolated polynucleotide having a polynucleotide sequence with a bZIP domain which is at least 25% identical to the bZIP domain of SEQ. ID. NO.: 5. 
     
     
         65 . The host cell of  claim 64 , wherein said B3-domain transcription factor is VP1 as defined by SEQ.ID.NO.:1. 
     
     
         66 . The host cell of  claim 65 , wherein said bZIP domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO. 5. and SEQ. ID. NO. 7. 
     
     
         67 . The host cell of  claim 64 , wherein said bZIP domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO. 5. and SEQ. ID. NO. 7. 
     
     
         68 . The host cell of  claim 64  wherein said bZIP domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO. 9, SEQ. ID. NO. 11, SEQ. ID. NO. 15, SEQ. ID. NO. 17, SEQ. ID. NO. 19, SEQ. ID. NO. 21, SEQ. ID. NO. 23, SEQ. ID. NO. 25 and SEQ. ID. NO. 27. 
     
     
         69 . The host cell of  claim 64  wherein said B3 domain transcription factor is selected from a group of polynucleotides consisting of SEQ. ID. NO. 29, SEQ. ID. NO. 31, SEQ. ID. NO. 33, SEQ. ID. NO. 35, SEQ. ID. NO. 37, SEQ. ID. NO. 39, SEQ. ID. NO. 41, SEQ. ID. NO. 43, SEQ. ID. NO. 45, SEQ. ID. NO. 47, SEQ. ID. NO. 49, SEQ. ID. NO. 51, SEQ. ID. NO. 53, SEQ. ID. NO. 55, SEQ. ID. NO. 57, SEQ. ID. NO. 59, SEQ. ID. NO. 61, SEQ. ID. NO. 63, and SEQ. ID. NO. 65. 
     
     
         70 . A vector comprising a first isolated polynucleotide having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID. NO. 1 and a second isolated polynucleotide having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID. NO.: 3. 
     
     
         71 . The vector of  claim 70 , wherein said first isolated polynucleotide is VP1 as defined by SEQ. ID. NO. 1. 
     
     
         72 . The vector of  claim 71 , wherein said second isolated polynucleotide is RAV2 as defined by SEQ. ID. NO.: 3. 
     
     
         73 . The vector of  claim 70 , wherein said second isolated polynucleotide is RAV2 as defined by SEQ. ID. NO.: 3. 
     
     
         74 . The vector of  claim 70 , wherein said first isolated polynucleotide or said second isolated polynucleotide is selected from a group of polynucleotides consisting of SEQ. ID. NO.: 29, SEQ. ID. NO.:31, SEQ. ID. NO.:33, SEQ. ID. NO.:35, SEQ. ID. NO.:37, SEQ. ID. NO.:39, SEQ. ID. NO.:41, SEQ. ID. NO.:43, SEQ. ID. NO.:45, SEQ. ID. NO.:47, SEQ. ID. NO.:49, SEQ. ID. NO.:51, SEQ. ID. NO.:53, SEQ. ID. NO.:55, SEQ. ID. NO.: 57, SEQ. ID. NO.:59, SEQ. ID. NO.:61, SEQ. ID. NO.:63, and SEQ. ID. NO.:65. 
     
     
         75 . A host cell comprising a first isolated polynucleotide having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID. NO. 1 and a second isolated polynucleotide having a polynucleotide sequence with a B3 domain which is at least 25% identical to the B3 domain of SEQ. ID. NO.: 3. 
     
     
         76 . The vector of  claim 75 , wherein said first isolated polynucleotide is VP1 as defined by SEQ. ID. NO. 1. 
     
     
         77 . The vector of  claim 76 , wherein said second isolated polynucleotide is RAV2 as defined by SEQ. ID. NO.: 3. 
     
     
         78 . The vector of  claim 75 , wherein said second isolated polynucleotide is RAV2 as defined by SEQ. ID. NO.: 3. 
     
     
         79 . The vector of  claim 75 , wherein said first isolated polynucleotide or said second isolated polynucleotide is selected from a group of polynucleotides consisting of SEQ. ID. NO.: 29, SEQ. ID. NO.:31, SEQ. ID. NO.:33, SEQ. ID. NO.:35, SEQ. ID. NO.:37, SEQ. ID. NO.:39, SEQ. ID. NO.:41, SEQ. ID. NO.:43, SEQ. ID. NO.:45, SEQ. ID. NO.:47, SEQ. ID. NO.:49, SEQ. ID. NO.:51, SEQ. ID. NO.:53, SEQ. ID. NO.:55, SEQ. ID. NO.: 57, SEQ. ID. NO.:59, SEQ. ID. NO.:61, SEQ. ID. NO.:63, and SEQ. ID. NO.:65.