Isolation and characterization of heat shock protein

Abstract

This invention relates to the identification and characterization of the maize HSP22 protein including the corresponding nucleic acid molecules, nucleic acid fragments and protein fragments. Further, this invention relates to both polyclonal and monoclonal antibodies to HSP22 and the production thereof. These antibodies can be used in a method for detecting the presence of HSP22 expression in whole leaf samples. Further, the nucleic acid molecule and fragments thereof can be used in assays to detect the levels of nucleic acid molecules involved in HSP22 expression. The levels of both HSP22 and HSP22 mRNA are then utilized as an indication of a plant's ability to tolerate heat stress. Additionally, plants can be transformed to express increased levels of HSP22.

We claim:  
     
         1 . A method for identifying individual entities of a plant species with an increased level of HSP22 during heat stress by comparing the respective quantities of HSP22 comprising: 
 a) collecting a batch of randomly selected individual entities of the plant species;    b) subjecting the plant species to heat stress; and    c) comparing the respective quantities of HSP22.    
     
     
         2 . The method claimed in  claim 1 , wherein the respective quantities of HSP22 are determined by utilizing an antibody to HSP22.  
     
     
         3 . The method claimed in  claim 2 , wherein the antibody to HSP22 is monoclonal.  
     
     
         4 . The method claimed in  claim 3 , wherein a fragment of the monoclonal selectively binds to HSP22.  
     
     
         5 . The method claimed in  claim 2 , wherein the antibody to HSP22 detects precursor HSP22.  
     
     
         6 . The method claimed in  claim 1 , wherein the plant species is maize.  
     
     
         7 . The method claimed in  claim 1 , wherein the respective quantities of HSP22 are determined by utilizing a nucleic acid probe that binds HSP22 mRNA.  
     
     
         8 . In combination: 
 at least one aliquoted substance that selectively binds to HSP22 or an HSP22 precursor to form a detectable combination; and    a detection method for detecting the combination.    
     
     
         9 . The combination as claimed in  claim 8 , wherein the aliquoted substance is an antibody.  
     
     
         10 . The combination as claimed in  claim 9 , wherein the antibody is monoclonal.  
     
     
         11 . The combination as claimed in  claim 9 , wherein the antibody is an antibody fragment.  
     
     
         12 . The combination as claimed in  claim 8 , wherein the aliquoted substance is a nucleic acid sequence.  
     
     
         13 . An in vitro method for determining a plant species ability to tolerate heat stress, comprising the step of determining if HSP22 is over expressed in a tissue sample, such over expression correlating directly with a plants ability to tolerate heat stress.  
     
     
         14 . A first isolated nucleic acid sequence that hybridizes to a second isolated nucleic acid sequence comprising SEQ. ID. NO. 7 under stringent conditions, and the first isolated nucleic acid sequence encodes a protein that has physical characteristics substantially equal to maize HSP22.  
     
     
         15 . A vector comprising a first nucleic acid sequence that hybridizes to a second nucleic acid sequence comprising SEQ. ID. NO. 7 under stringent conditions, and the first nucleic acid sequence encodes a protein that has physical characteristics substantially equal to HSP22.  
     
     
         16 . The vector of  claim 15 , wherein the vector contains regulatory elements operably linked to the nucleic acid sequence.  
     
     
         17 . A plant transformed with the vector of  claim 15 .  
     
     
         18 . The plant of  claim 17  wherein the plant is a dicot.  
     
     
         19 . The plant of  claim 17  wherein the plant is a monocot.  
     
     
         20 . The plant of  claim 17  wherein the plant is maize.  
     
     
         21 . The plant of  claim 17  where in the plant is Arabidopsis.  
     
     
         22 . A host cell transformed with a vector comprising a first nucleic acid sequence that hybridizes to a second nucleic acid sequence comprising SEQ. ID. NO. 7 under stringent conditions, and the first nucleic acid sequence encodes a protein that has physical characteristics substantially equal to maize HSP22.  
     
     
         23 . The host cell of  claim 22  wherein the host cell is prokaryotic.  
     
     
         24 . The host cell of  claim 22  wherein the host cell is eukaryotic.  
     
     
         25 . Isolated protein encoded by a first nucleic acid sequence that hybridizes to a second nucleic acid sequence comprising SEQ. ID. NO. 7 under stringent conditions, and the first nucleic acid sequence encodes a protein that has physical characteristics substantially equal to maize HSP22.  
     
     
         26 . A method for the production of an isolated protein encoded by a first nucleic acid sequence that hybridizes to a second nucleic acid sequence comprising SEQ. ID. NO. 7 under stringent conditions, and the first nucleic acid sequence encodes a protein that has physical characteristics substantially equal to maize HSP22 comprising the steps of: 
 a) causing a host cell to produce the protein; and    b) isolating the protein.    
     
     
         27 . A method for the production of isolated nucleic acid sequences that hybridize to a nucleic acid sequence comprising SEQ. ID. NO. 7 under stringent conditions, and the isolated nucleic acid sequences encode a protein that has physical characteristics substantially equal to maize HSP22 comprising the steps of: 
 a) causing a host cell to produce the nucleic acid sequence; and    b) isolating the nucleic acid sequence.    
     
     
         28 . The vector of  claim 15  wherein the vector is capable of expressing HSP22.  
     
     
         29 . An isolated nucleic acid sequence encoding maize HSP22.  
     
     
         30 . A nucleic acid sequence comprising SEQ. ID. NO. 1.  
     
     
         31 . A nucleic acid sequence comprising SEQ. ID. NO. 4.  
     
     
         32 . A nucleic acid sequence comprising SEQ. ID. NO. 5.  
     
     
         33 . A nucleic acid sequence comprising SEQ. ID. NO. 7.  
     
     
         34 . A first isolated nucleic acid sequence that hybridizes to a second isolated nucleic acid sequence comprising SEQ. ID. NO. 1 under stringent conditions, and the first isolated nucleic acid sequence encodes a protein that has physical characteristics substantially equal to maize HSP22.  
     
     
         35 . A first isolated nucleic acid sequence that hybridizes to a second isolated nucleic acid sequence comprising SEQ. ID. NO. 4 under stringent conditions, and the first isolated nucleic acid sequence encodes a protein that has physical characteristics substantially equal to maize HSP22.  
     
     
         36 . A first isolated nucleic acid sequence that hybridizes to a second isolated nucleic acid sequence comprising SEQ. ID. NO. 5 under stringent conditions, and the first isolated nucleic acid sequence encodes a protein that has physical characteristics substantially equal to maize HSP22.  
     
     
         37 . An antibody that selectively binds to a portion of HSP22.  
     
     
         38 . The antibody of  claim 37  wherein the antibody is polyclonal.  
     
     
         39 . The antibody of  claim 37  wherein the antibody is monoclonal.  
     
     
         40 . The antibody of  claim 37  wherein HSP22 is from maize.  
     
     
         41 . A hybridoma cell line that produces a monoclonal antibody that selectively binds to HSP22.  
     
     
         42 . The antibody of  claim 37  wherein the antibody is directed against at least a portion of SEQ. ID. NO. 2.  
     
     
         43 . The antibody of  claim 37  wherein a fragment of the antibody will selectively bind HSP22.  
     
     
         44 . A process for producing a monoclonal antibody that selectively binds to a portion of HSP22 comprising the steps of: 
 a) culturing a hybridoma that produces the monoclonal antibody; and    b) isolating the monoclonal antibody.    
     
     
         45 . An isolated protein comprising SEQ. ID. NO. 6.  
     
     
         46 . An isolated nucleic acid sequence that encodes a protein comprising SEQ. ID. NO.6.  
     
     
         47 . A vector comprising a first nucleic acid sequence that hybridizes to a second nucleic acid sequence comprising SEQ. ID. NO. 5 under stringent conditions, and the first nucleic acid sequence encodes a protein that has physical characteristics substantially equal to HSP22.  
     
     
         48 . The vector of  claim 47 , wherein the vector contains regulatory elements operably linked to the nucleic acid sequence.  
     
     
         49 . A plant transformed with the vector of  claim 47 .  
     
     
         50 . The plant of  claim 49  wherein the plant is a dicot.  
     
     
         51 . The plant of  claim 49  wherein the plant is maize.  
     
     
         52 . The plant of  claim 49  wherein the plant is Arabidopsis.  
     
     
         53 . A plant species modified to express an amount of HSP22 in excess of a naturally occurring amount of HSP22.