关键词: 瘤胃微生物, BAC文库, 乙酰CoA羧化酶, 基因注释, 生物信息学

Abstract:

【Objective】 The objective of the experiment is to screen, sequence and bioinformatical analyze the ACCase clone from a BAC library of the dairy cow rumen microbiota. 【Method】 ACCase gene was screened from BAC library of the rumen microbiota by PCR sequence-driven analysis. Whole insert sequence of screened clone was sequenced by shotgun method. Then it was analyzed by gene annotation, blast and phylogenetic tree analysis. 【Result】 One ACCase clone (U12) was obtained , and its insert fragment (URE12) had the length of 43 358 bp and GC content of 43.75%. URE12 contained 38 CDSs predicted by GeneMark software, and it was probably originated from Proteobacteria by Signature analysis. ACCase encoded by Acc-32 was predicted to be 159 amino acids with a molecular weight of 17.89 kD and pI of 5.27. Acc-32 had the similarity of 41% with that from Elusimicrobium minutum. The combining site AA residue of Acc-32 was GQVICVIEAMKVFNELKA, and Acc-32 belonged to ε-Proteobacteria by phylogenetic tree analysis. 【Conclusion】 The fragment containing ACCase was screened, and ACCase had some novel gene characteristics.

Key words: rumen microbiota, BAC library, acetyl-CoA carboxylase, gene annotation, bioinformatics analysis