Po. Sheppard et al., THE USE OF CONSERVED CELLULASE FAMILY-SPECIFIC SEQUENCES TO CLONE CELLULASE HOMOLOG CDNAS FROM FUSARIUM-OXYSPORUM, Gene, 150(1), 1994, pp. 163-167
Five cDNAs from the cellulolytic fungi Fusarium oxysporum that code fo
r five distinct cellulase homologues have been cloned and sequenced. T
he cloning strategy exploited the hydrophobic cluster analysis-based c
ellulase family classification of Henrissat and Bairoch [Biochem. J. 2
93 (1993) 781-788] to design degenerate oligodeoxyribonucleotides (oli
gos) that encoded amino-acid sequences conserved in an intra-family, b
ut not inter-family, manner among cellulases from different species. P
olymerase chain reaction (PCR) experiments using F. oxysporum genomic
DNA primed with these 'family-specific' oligos were used to rapidly ge
nerate PCR fragments which were in turn used to probe cDNA libraries.
Two distinct cDNAs coding for cellulase C-family homologues and one cD
NA each coding for homologues to the B, F and K families, were isolate
d in this manner. This approach is an example of the power of multiple
sequence analysis to generate cross-species, homology-based probes to
rapidly clone homologues in a species of interest.