Open AccessMolecular cloning of mRNA sequences encoding xylanase from Cryptococcus albidus
Author(s) -
Morosoli Rolf,
Durand Serge
Publication year - 1988
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1988.tb03000.x
Subject(s) - complementary dna , biology , microbiology and biotechnology , cdna library , molecular cloning , plasmid , coding region , nucleic acid sequence , xylanase , gene , sequence analysis , genetics , biochemistry , enzyme
A cDNA library has been prepared from Cryptococcus albidus mRNA using the plasmid expression vector pUC13. Hin dIII linkers were added to the cDNA ends and the cDNA was inserted into the Hin dIII‐cut plasmid vector, pUC13, which contains the transcriptional and translational start sequences of the lac Z gene. The colonies were replica plated onto nitrocellulose filters and screened by differential colony hybridization using 32 P‐labeled cDNA synthesized from mRNA isolated from cells grown on xylan and xylose. The cDNA clones identified were further analyzed by hybrid‐selected translation. A 540 bp cDNA clone (pIAF101) was shown to contain a part of the xylanase sequence, and this was confirmed by DNA sequence analysis, by comparing the coding sequence with the partial amino acid sequence of the xylanase.