Chukeatirote Ekachai, Maharachchikumbura Sajeewa S N, Wongkham Shannaphimon, Sysouphanthong Phongeun, Phookamsak Rungtiwa, Hyde Kevin D
Institute of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand.
Mycobiology. 2012 Jun;40(2):107-10. doi: 10.5941/MYCO.2012.40.2.107. Epub 2012 Jun 29.
Genes encoding the cellobiohydrolase enzyme (CBHI), designated as cbhI, were isolated from the basidiomycetes Auricularia fuscosuccinea, Pleurotus giganteus, P. eryngii, P. ostreatus, and P. sajor-caju. Initially, the fungal genomic DNA was extracted using a modified cetyltrimethyl ammonium bromide (CTAB) protocol and used as a DNA template. The cbhI genes were then amplified and cloned using the pGEM-T Easy Vector Systems. The sizes of these PCR amplicons were between 700~800 bp. The DNA sequences obtained were similar showing high identity to the cbhI gene family. These cbhI genes were partial consisting of three coding regions and two introns. The deduced amino acid sequences exhibited significant similarity to those of fungal CBHI enzymes belonging to glycosyl hydrolase family 7.
编码纤维二糖水解酶(CBHI)的基因,命名为cbhI,是从担子菌黑木耳、巨大侧耳、刺芹侧耳、糙皮侧耳和平菇中分离得到的。最初,使用改良的十六烷基三甲基溴化铵(CTAB)方法提取真菌基因组DNA,并将其用作DNA模板。然后使用pGEM-T Easy载体系统对cbhI基因进行扩增和克隆。这些PCR扩增产物的大小在700至800 bp之间。获得的DNA序列相似,与cbhI基因家族具有高度同源性。这些cbhI基因是部分基因,由三个编码区和两个内含子组成。推导的氨基酸序列与属于糖基水解酶家族7的真菌CBHI酶的氨基酸序列具有显著相似性。