Molecular biology tools for algal engineering

Abstract

The present invention provides compositions and methods for the genetic manipulation of Algal cells. The compositions and methods allow enhanced transfer of genetic material into Algal cells and the cloning and selection of genetically modified cells. Expression of proteins encoded by the genetic material will be enhanced by the methods and compositions of the invention.

1 . A composition for transformation of an Algal cell with DNA comprising:
 (a) one or more sugars, and   (b) a biological buffer.   
     
     
         2 . The composition of  claim 1 , wherein the one or more sugars are selected from the group consisting of sucrose, fructose, maltose, trehalose, sorbitol, maltitol, erythrytol, mannitol, xylose, raffilose and lactose. 
     
     
         3 . The composition of  claim 2 , wherein the one or more sugars are sucrose and sorbitol. 
     
     
         4 . The composition of  claim 1 , wherein the one or more sugars are present at a total concentration of from 40 mM to 100 mM. 
     
     
         5 . The composition of  claim 1 , wherein the biological buffer is selected from the group consisting of Bis-Tris Propane, TRIS, AMPD, TABS, AMPSO, CHES and CAPSO. 
     
     
         6 . The composition of  claim 1 , wherein the concentration of the biological buffer is from 5 mM to 100 mM. 
     
     
         7 . The composition of  claim 1 , wherein the pH is from 8 to 10. 
     
     
         8 . The composition of  claim 7 , wherein the pH is from 9 to 10. 
     
     
         9 . The composition of  claim 1 , further comprising an algal cell. 
     
     
         10 . The composition of  claim 9 , wherein the algal cells is of the genus  Chlorella  or  Chlamydomonas.    
     
     
         11 . A method for transformation of DNA into an algal cell comprising:
 (a) suspending the algal cells in a buffer,   (b) adding DNA to be transformed to the algal cell suspension,   (c) applying an electric pulse to the DNA Algal cell suspension thereby transforming the algal cell with the DNA.   
     
     
         12 . The method of  claim 11 , wherein the buffer comprises one or more sugars and a buffer salt. 
     
     
         13 . The method of  claim 12 , wherein the one or more sugars are selected from the group consisting of sucrose, fructose, maltose, trehalose, sorbitol, maltitol, erythrytol, mannitol, xylose, raffilose and lactose. 
     
     
         14 . The method of  claim 12 , wherein the one or more sugars are sucrose and sorbitol. 
     
     
         15 . The method of  11 , wherein the pH of the buffer is from pH 8 to pH 10. 
     
     
         16 . The method of  claim 15 , wherein the pH of the buffer is from pH 9 to pH 10. 
     
     
         17 . A kit for the transformation of DNA into an Algal cell comprising:
 (a) a buffer,   (b) algal cells.   
     
     
         18 . The kit of  claim 17 , wherein the one or more sugars are selected from the group consisting of sucrose, fructose, maltose, trehalose, sorbitol, maltitol, erythrytol, mannitol, xylose, raffilose and lactose and a buffer salt selected from the group consisting of Bis-Tris Propane, TRIS, AMPD, TABS, AMPSO, CHES and CAPSO. 
     
     
         19 . The kit of  claim 18 , wherein the pH of the buffer is from pH 8 to pH 10. 
     
     
         20 . The kit of  claim 18 , further comprising and electroporation cuvette. 
     
     
         21 - 27 . (canceled)