Trigui-Lahiani Hèla, Abdeljalil Salma, Hadj Sassi Azza, Skouri-Gargouri Houda, Gargouri Ali
Laboratory of Molecular Biotechnology of Eukaryotes, Centre of Biotechnology of Sfax, P.O. Box 1177, 3038 Sfax, University of Sfax, Tunisia.
Laboratory of Molecular Biotechnology of Eukaryotes, Centre of Biotechnology of Sfax, P.O. Box 1177, 3038 Sfax, University of Sfax, Tunisia.
Genomics. 2021 Jan;113(1 Pt 2):795-811. doi: 10.1016/j.ygeno.2020.10.011. Epub 2020 Oct 16.
As a preliminary step to characterize genes encoding ATP-Binding-Cassette (ABC) proteins, we cloned a gene encoding an ABC transporter from P. occitanis using a PCR based approach followed by a genomic library screening and by additionally using whole genome sequencing results. The encoded protein has high similarity to the pleiotropic drug resistance protein subfamily members. Analysis of the cloned sequence revealed the presence of Walker A, Walker B and the ABC signature motifs at the nucleotide binding domains. Molecular docking resulted in predicting the most stable complex between the gene-encoding protein and cycloheximide. The southern blot results indicate that the gene is present as a single copy in the P. occitanis genome. The genome-scale identification of the PoABC superfamily members led to the characterization of 58 putative proteins divided into five subfamilies including: 12 ABCB, 24 ABCC, 1 ABCE, 5 ABCF, 15 ABCG, and of which 51 contain trans-membrane domains.
作为表征编码ATP结合盒(ABC)蛋白的基因的初步步骤,我们采用基于PCR的方法,随后进行基因组文库筛选,并额外利用全基因组测序结果,从奥克西坦尼假丝酵母中克隆了一个编码ABC转运蛋白的基因。所编码的蛋白质与多药耐药蛋白亚家族成员具有高度相似性。对克隆序列的分析揭示了在核苷酸结合结构域存在沃克A、沃克B和ABC特征基序。分子对接预测了编码该基因的蛋白质与环己酰亚胺之间最稳定的复合物。Southern杂交结果表明该基因在奥克西坦尼假丝酵母基因组中以单拷贝形式存在。对PoABC超家族成员的全基因组鉴定导致了58个推定蛋白的表征,这些蛋白分为五个亚家族,包括:12个ABCB、24个ABCC、1个ABCE、5个ABCF、15个ABCG,其中51个含有跨膜结构域。