首页|期刊导航|中国农业科学（英文版）|Isolation, Cloning, and Identification of Expressed Sequence Tags from the Backfat Tissue in Duroc and Tongcheng Pigs by mRNA Differential Display
中国农业科学（英文版）2006，Vol.5Issue(2)：141-145,5.
Isolation, Cloning, and Identification of Expressed Sequence Tags from the Backfat Tissue in Duroc and Tongcheng Pigs by mRNA Differential Display
Isolation, Cloning, and Identification of Expressed Sequence Tags from the Backfat Tissue in Duroc and Tongcheng Pigs by mRNA Differential Display
摘要
Abstract
mRNA differential display technique was performed to discuss the differential expression of genes in fat tissue between introduced European and Chinese indigenous pigs. Four anchor primers in combination with five arbitrary primers (20 sets in total) were used and nearly 300 bands were observed in polyacrylamide gel, among which 29 differential display bands were obtained. Twelve of 29 eDNA fragments were identified using reverse Northern dot blot, and subsequently cloned and sequenced. Eight of 12 cDNAs had no matches in GenBank and were submitted to GenBank, and the other 4 showed similarity to identified genes from GenBank. Three among 8 novel ESTs were selected to be further identified by semiquantitative RT-PCR. In our experiment, silver staining DDRT-PCR and DIG primer DNA labeling reverse Northern dot blot were used to avoid radioactive pollution. The result showed that the expressions of 5 among 8 novel ESTs were stronger in the backfat of Tongcheng pigs and the others were weaker than that in Duroc pigs. These novel ESTs were prepared for selecting genes related to adipose cells.关键词
ESTs/fat tissue/reverse Northern dot blot/semi-quantitative RT-PCR/mRNA differential displayKey words
ESTs/fat tissue/reverse Northern dot blot/semi-quantitative RT-PCR/mRNA differential display分类
农业科技引用本文复制引用
REN Zhu-qing,XIONG Yuan-zhu,DENG Chang-yan,LEI Ming-gang,ZUO Bo,LI Feng-e,ZHENG Rong ,XU De-quan..Isolation, Cloning, and Identification of Expressed Sequence Tags from the Backfat Tissue in Duroc and Tongcheng Pigs by mRNA Differential Display[J].中国农业科学（英文版）,2006,5(2):141-145,5.基金项目
This study was supported by the National 973 Project of China (G2000016105). （G2000016105）
