Abstract:

【Objective】 This study was carried out to construct suppression subtractive hybridization(SSH) library of secondary follicle in anagen and to search some candidate genes involved in cashmere development in Inner Mongolia cashmere goat. 【Method】 cDNA subtracted library was performed with cDNA from secondary follicle in anagen as the “tester” and cDNA from secondary follicle in telogen as the “driver”. The gene fragments were sequenced and analyzed by bioinformatics. 【Result】 The cDNA library was constructed successfully. Twenty positive clones were amplified by using nested PCR primer 1 and 2R, the size of inserts was 250-1 000 bp. Three hundred and fourty-two genes were obtained by DNA sequencing from 750 positive clones picked randomly. Their average length is 596 bp. After nucleotide blast homological analysis, 298 matched to known genes, 38 matched only to other ESTs in dbEST, and the remaining six showed no match to any ESTs or known genes. A total of 298 unique known genes were used to analyze the gene expression patterns. These were categorized into seven categories on the basis of gene function. They were divided into cell division (13), cell signaling/communication (49), cell structure/motility (52), cell/organism defense (21), metabolism (46), gene/protein expression (37), and unclassified (80). 【Conclusion】 The subtractive cDNA library of secondary follicles in anagen and telogen were obtained by SSH, and the information generated in this study had established a basis for screening candidate genes involved in cashmere development.

Key words: cashmere goat, secondary follicle, anagen, telogen, suppression subtractive hybridization, sequence analysis