Glycosyltransferase2 gene silD in E. coli Cloning and Expression of.docVIP

 PAGE \* MERGEFORMAT 17 Glycosyltransferase2 gene silD in E. coli Cloning and Expression of Author: Wen-Rong Zeng Zhi-hong ZHU Chun-bao, CHEN Dai-Jie ZHU Bao-Quan [Abstract] From the Yiniu Micromonospora strain (Micromonospora inyoensis) amplification involved in the biosynthesis of sisomicin glycosyltransferase gene silD (1181bp), respectively, cloned into pUC18 and the expression vector pET  30a. Its open reading frame of 1173bp, encoding with 390aa (43.04ku) peptide chains in E. coli BL21 (DE3):: pET  silD to achieve expression. The base sequence of genes silD and gntD (from M.echinospora) nucleotide sequence homology as high as 94%. Forecast SilD protein sequence homology with the GntD of 98%. This is in accordance with New Zealand, from the cloning of small single-spore bacteria involved in the biosynthesis of sisomicin resistance genes srm1 (AY661430), sisomicin 2  deoxy-myo-inositol synthase gene crab sisB (DQ250993) and the glycosyl transferase gene sisZ (DQ250994), the new gene cloned into another. The acceptance of the gene in GenBank No. DQ250992. SilD cloning and expression of genes for further study on the basis of its biological function. [Keywords:] Glycosyltransferase  2 gene; Yiniu Micromonospora strain; silD; Cloning and Expression of ABSTRACT The gene silD, a novel glycosyl transferase  2 gene, was cloned in our laboratory. The DNA fragment containing silD (1185bp) gene (the key gene involved in sisomicin biosynthesis) was isolated from Micromonospora inyoensis and cloned into pUC18 vector. Its ORF is composed of 1,173 bp, encoding 390aa (43.04ku). Sequence analysis verified that silD gene and amino acids sequence showed 94% and 98% identity with gntD (glycosyltransferase gene for gentamicin biosynthesis from M.echinospora), respectively. The prediction of structure of silD suggested it is also a glycosyl transferase protein. The silD expression vector pET  silD was constructed by fusion of the flanking silD ORF in the pET 