Hypoallergenic transgenic soybeans

Abstract

Hypoallergenic transgenic soybeans and recombinant expression constructs to lower soybean vacuolar protein, commonly know as P34, as well as other allergens are disclosed. Soybean protein products made from these hypoallergenic soybeans should be substantially free of the major soy allergen, P34, and, in addition, other minor soy allergens, such as, Gly m Bd 28K, alpha-subunit of beta-conglycinin, KSTI, Gly m2, Gly m IA, Gly m IB, rGLY m3 and glycinin G1.

1 . A recombinant expression construct to lower Gly m Bd 30K (Soybean vacuolar protein P34) content of a soybean which comprises a promoter operably linked to an isolated Gly m Bd 30K nucleic acid fragment corresponding substantially to the nucleotide sequence set forth in SEQ ID NO:1 or a functionally equivalent subfragment thereof.  
     
     
         2 - 90 . (canceled)  
     
     
         91 . A method for making a hypoallergenic soybean plant which comprises: 
 (a) crossing a (i) first parent soybean which is a soybean plant comprising in its genome a recombinant expression construct to lower the Gly m Bd 30K (Soybean vacuolar protein P34) content of a soybean which comprises a beta-conglycinin promoter operably linked to an isolated Gly m Bd 30K nucleic acid fragment corresponding to all or a part of the nucleotide sequence set forth in SEQ ID NO: 1, with a (ii) second parent soybean which is a hybrid, mutant or transgenic soybean parent wherein said second parent is substantially free of one or more allergens selected from the group consisting of Gly m Bd 28K, alpha-subunit of beta-conglycinin, KSTI, Gly m2, Gly m IA, Gly m IB, rGLY m3 and Glycinin G1; and    (b) selecting progeny plants of the cross of step (a) which are hypoallergenic.    
     
     
         92 . A method for making a hypoallergenic soybean plant which comprises: 
 (a) crossing a (1) first parent soybean which is a soybean plant comprising in its genome at least one expression construct selected from the group consisting of: 
 (i) a recombinant expression construct to lower Gly m Bd 30K (Soybean vacuolar protein P34) content of a soybean which comprises a promoter operably linked to an isolated Gly m Bd 30K nucleic acid fragment corresponding to all or a part of to the nucleotide sequence set forth in SEQ ID NO:1;  
 (ii) the recombinant expression construct of part (a)(i) wherein the promoter is selected from the group consisting of an alpha-subunit beta-conglycinin promoter, a Kunitz Trypsin Inhibitor (KSTI) promoter, a Gly m Bd 28K promoter, T7 promoter, a  35 S promoter and a beta-phaseolin promoter;  
 (iii) the expression construct of  claim 91;  and  
 (iv) a recombinant expression construct for producing a hypoallergenic soybean which comprises an isolated KSTI nucleic acid fragment corresponding to all or part of the nucleotide sequence set forth in SEQ ID NO:2 operably linked to an isolated Gly m Bd 28K nucleic acid fragment corresponding to all or part of the nucleotide sequence set forth in SEQ ID NO:3 with a  
 (2) second parent soybean which is a hybrid, mutant or transgenic soybean wherein the second parent is selected from the group consisting of a soybean plant which is substantially free of the alpha subunit of beta-conglycinin; and  
   (b) selecting progeny plants of the cross of step (a) which are hypoallergenic.    
     
     
         93 . A method for making a hypoallergenic soybean plant which comprises: 
 (a) crossing a (i) first parent soybean which is a soybean plant comprising in its genome at least one of the expression constructs of  claim 92  with a (ii) second parent soybean which is a hybrid, mutant or transgenic soybean wherein the second parent is substantially free of KSTI allergen; and    (b) selecting progeny plants of the cross of step (a) which are hypoallergenic.    
     
     
         94 . A method for making a hypoallergenic soybean plant which comprises: 
 (a) crossing a (i) first parent soybean which is a soybean plant comprising in its genome at least one of the expression constructs of  claim 92  with a (ii) second parent soybean which is a hybrid, mutant or transgenic parent soybean wherein said second parent is substantially free of soybean allergens selected from the group consisting of Gly m Bd 28K, alpha-subunit of beta-conglycinin, KSTI, Gly m2, Gly m IA, Gly m IB, rGLY m3 and Glycinin G1; and    (b) selecting progeny plants of the cross to step (a) which are hypoallergenic.    
     
     
         95 . Seeds obtained from a plant made by the method of any of claims  91 ,  92 ,  93  or  94 .