Cutinase cloning

The structure of the cutinase gene has been determined using genomic DNA cloned in X-phage. This gene contains one 51 bp intron which has the typical junction and splicing signals... 

Figure 2. Nucleotide sequence of the cloned cutinase cDNA and the amino acid sequence deduced from it, C-4 and C-57 indicate the beginning of the nucleotide sequence of two additional cDNA clones. The solid lines represent the regions for which the primary structure was confirmed by amino acid sequencing.

Figure 3 Amino acid sequences of cutinases from a Colletotrichum and a Fusarium species deduced from the nucleotide sequence of cloned cDNA. Regions of complete homology are shown by the lines. The aspartic acid, serine and histidine residues indicated by the arrows are the members of the catalytic triad and the position of the cys residues are also indicated by arrows. The two cys residues closest to the carboxyl terminus are in disulphide bridge with each other and so are the other two cys residues. The positions of the intron which interrupts the coding sequences in the genes are indicated by the vertical lines.

Figure k The nucleotide sequence of cutinase gene from Fusarium solani pisi. Southern blots show that this isolate contains another copy of the gene. However, the copy shown above is completely homologous with the cloned cDNA which would code for a protein with the primary structure that matches the major cutinase isozyme secreted by the fungus. 

I98A) Cloning and structure determination of cDNA for cutinase, an enzyme involved in fungal penetration of plants. Proc. Natl. Acad. Sci. USA. 81, 3939-39 3. 

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