| 摘要 |
<p>A method of cloning of cellulase cDNA clones with enzymatical activity from an anaerobic rumen fungus including the steps of: (i) cultivation of an anaerobic rumen fungus; (ii) isolating total RNA from the culture in step (i); (iii) isolating poly A+ mRNA from the total RNA referred to in step (ii); (iv) constructing a cDNA expression library; (v) ligating cDNA to a bacteriophage expression vector selected from μZAP, μZAPII or vectors of similar properties; (vi) screening of cellulase positive recombinant clones in a culture medium incorporating cellulose by detection of cellulose hydrolysis; and (vii) purifying cellulase positive recombinant clones. There is also provided recombinant cellulase fungal cDNA clones produced by the abovementioned method as well as the recombinant cellulase cDNA clones, derived from N. patriciarum, having the property of production of biologically functional cellulases in E. coli cells. There is also provided various cDNA molecules which may be utilised in the abovementioned method.</p> |
