Cloning and expression analyses of a cellobiohydrolase gene from Auricularia heimuer. Issue 1 (1st January 2019)

• Record Type: Journal Article
• Authors: Sun, Jian; Wang, Shixin; Wang, Xutong; Sun, Tingting; Zou, Li
• Is Part Of: Biotechnology, biotechnological equipment. Volume 33:Issue 1(2019)
• Page Start: 1327
• Page End: 1334
• Publication Date: 2019-01-01
• Subjects: Auricularia heimuer -- cellobiohydrolase -- prokaryotic expression -- quantitative real-time PCR; Biotechnology -- Periodicals; Biotechnology -- Periodicals; Biotechnology -- instrumentation -- Periodicals; Periodicals; 660.605
• DOI: 10.1080/13102818.2019.1665477
• Languages: English
• ISSNs: 1310-2818

Abstract

Abstract: A cellobiohydrolase gene was cloned and analyzed from Auricularia heimuer, and named A-cbh . The cDNA sequence was submitted to GenBank under the accession number MK620906. Sequence analysis showed that the cDNA comprised 1551 bp and encoded a putative polypeptide of 516 amino acids. The molecular weight of this protein was predicted as 54.44 kDa, and the theoretical isoelectric point was 5.24. The recombinant prokaryotic expression vector pET32a- A-cbh was constructed and transformed into Escherichia coli BL21. Sodium dodecyl sulphate polyacrylamide gel electrophoresis indicated that the recombinant protein had molecular size between 66.2 kDa and 94.0 kDa, which was consistent with our expectations because a tag protein of approximately 20 kDa was present in the vector of pET32a. The transcription levels of A-cbh in the mycelium were lower than those in the fruiting bodies. The highest level of A-cbh expression was observed in the primordium, which was 8.65-fold higher than that in the control (mycelia at 6 days). We speculated that A-cbh might be involved in the formation of primordia and the development of fruiting bodies. The results of the present study will form the basis for further research into the function of the cellobiohydrolase in A. heimuer .