Cloning and Sequence Analysis of Cellulose Synthase-like Gene from Ampelopsis megalophylla / 中国实验方剂学杂志

ABSTRACT

Objective:To clone cellulose synthase-like(Csl)gene from ethnic medicinal plant Ampelopsis megalophylla,and analyze its sequence by bioinformatics. Method:Specific primers were designed for AmCsl gene sequences obtained from A. megalophylla transcriptome sequencing data. The full-length cDNA of AmCsl gene was amplified by PCR using cDNA of young leaves as template,and TA clone and sequencing was performed. The sequence was analyzed by bioinformatics. Result:The full length cDNA was 1 438 bp,containing a 561 bp open reading frame(ORF),and encoding 186 amino acids,the molecular formula of protein was C1011H1547N233O257S10,the theoretical relative molecular weight was 22.40 kDa,the theory isoelectric point(PI)was 7.59,and the aliphatic index(AI)was 116.88.There was a transmembrane region and no signal peptide,which may be located in the endoplasmic reticulum,the average hydrophobic coefficient was 0.670,and the instability index was 42.56.It belonged to a hydrophobic unstable protein. The conserved domain contained a cellulose synthase,and the secondary structure mainly was dominated by α-helix. Amino acid sequence alignment and phylogenetic tree analysis showed that AmCsl had a high homology with Vitis vinifera. Conclusion:The full length of AmCsl gene was obtained for preliminary bioinformatics analysis,which laid a necessary foundation for further study on the accumulation of polysaccharides and the regulation of biosynthesis.