In silico Cloning of Xyloglucan Endotransglycosylase( XET) GeneRelated to Fertility Changeover in the Thermo-sensitive ChineseCabbage(Brassica campestris L. ssp. pekinensis)
1College of Horticulture, Northwest Agricultural and Forestry University, Yangling 712100, Shaanxi, China;2Vegetables Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, Gansu, China
Abstract:The EST-58 related to the fertility changeover in the thermo-sensitive CMS
(TsCMS7311)of Chinese cabbage(Brassica campestris L. ssp. pekinensis)was amplified by in silico cloning. And the cDNA sequence had been confirmed by RT-PCR. The result indicated that a new sequence with 1 197 bp length was cloned,which contained an integrated ORF, encoding 292 amino acids.
The trend of EST-58 expression by RT-PCR was very identity to the expression by cDNA-AFLP, and their sequences have 98 % consistency. As showed that the result of in silico cloning was right. The amino acid sequence showed highly comparability with xyloglucan endotransglycosylase(XET)from GenBank. It
indicated that the new sequence was XET gene(GenBank accession,EU579461)of Chinese cabbage.