Construction Of The Suppression Subtractive Hybridization Library Of Hybrid Argali Tail And Preliminary Analysis Of The Transcriptome

In sheep industries the lamb’s tail need to cut by workers. Not only increase the workload and improper operation will make the lamb died. Our sheep farm has 75% hybrid argali sheep, its tail is short. In order to investigate the short tail formation mechanism of argali sheep,suppression subtractive hybridization(SSH) and RNA-Seq technologies were used to study the 25% and 75% hybrid argali tails. And the subtracted library and different m RNAs were obtained to find the tail developing related functional genes and to breed new short tail sheep. We used the 25% hybrid argali sheep tail as the experimental material, and the 75% hybrid argali sheep tail as control to construct two forwad- and reverse-substracted c DNA libraries. In the two libraries, 200 clones of each library were sent to company to make sequencing. There were 129 genes in forwad-substracted c DNA libraries and 109 genes in reverse-substracted were successfully sequenced. Genetic variations are randomly selected from within the library the 44, the reverse random took 15 different genes.As a result, positive library FN1 TBX2 and LGALS3 and reverse library with TBX2 TBX18 may be associated with skeleton.The Hiseq2000 platform was also used for high-throughput transcriptome sequencing, then the results analysis were processed through blasting in NR, GO and KEGG, making gene comments and clustering analysis to detect differentially expressed genes.129 and 106 ESTs were identified from positive and negative c DNA library. The high-throughput transcriptome sequencing show that there were 133542 unigenes in 25% and 136318 unigenes in 75% hybrid argali tail. The GO function classification annotations to the biological process, the molecular and cellular composition in the database with 195878, 195878 and 26200 respectively. Annotation to KEGG database, the number of all the genes in KEGG pathways is 36503, including 4807 differentially expressed genes. The total number differentially expressed genes of the argali sheep is 8515.The 25% has 3474 genes up-regulation and 5041 genes Down-regulation reference to 75% hybrid argali. Rise in the three largest gene is TPM3, MYH1 and TNNT1, one of the biggest declines genes is MHC(-15.3141), RNASET2(-14.3305)and KRTAP10-8(-13.9062). The transcription sequencing had found T-box family genes(TBX2 TBX3 TBX15 TBX18 and TBX19), including TBX3, TBX15 and TBX18 about bone growth and development, is likely to play a key role in small tail formation mechanism.Finally we selected TBX18 and TBX15 as potential target genes of sheep with small tail traits.