Construction Of Normalized Full-length CDNA Library Of Xinji Fine-wool Sheep Skin Tissue And Bioinformatics Analysis Of Its ESTs

Xinji Fine-wool Sheep is the first super-fine-wool breed cutivited in China, its wool fiber fitness is between 66 and 70 s(a few up to 80s), and wool length is above 8 cm. It is the ideal material for slap-up textile. Acccouting for the features of Xinji Fine-wool Sheep, a main aspect on how to maintain and keep cutiviting more fine-wool breeds is to clone and idendify some genes related to the expression of wool trait, then study on the fuctions of those genes and finally apply to productive practical of Xinji Fine-wool Sheep. Combining bioinformatics with biotechnology, a set of new genes were cloned and identified. The main results are as follows:1. Normalized Full-length cDNA library of Xinji Fine-wool Sheep skin tissue was constructed by using CreaterTM SMARTTM cDNA Library Construction Kit and DSN enzyme. The results showed that the titer of the library is 1.1×l06,the recombinant percentage is 99%,the average inserts length is about 1.2 kb.2. 75 EST sequences are obtained from 5'end randomly single-pass sequencing of full-length cDNA library of Xinji Fine-wool Sheep skin tissue and assembled to 4 Contigs and 67 Singletons by SeqMan Program of DNAStar soft. Blastn Results showed that 10 ESTs may be known and the rest are unknown. The results of full length cDNA annotation showed that 6 EST sequences may contain intact gene and another 6 may have complete CDS.3. 25 of 49 EST sequences which did not have intact gene or complete CDS were effectively electro-extended. The results of full length cDNA annotation showed that 3 EST sequences may contain intact genes, and 4 EST sequences may contain complete CDS.4. The nucleotide sequences of 9 EST sequences covering the whole genes were predicted and their basic properties, electronic expressed profiles were analyzed. 19 ESTs which contains whole genes and complete CDS were translated into proteins, and basic properties, structures and functions of them were analyzed, too. The results indicated that 19 EST sequences are more or less containing some important functional sites and fuctional structure domains.