Transcriptome Analysis Of Roe Deer Antler Tip And Functional Genes Cloning Detection

Roe deer (Capreolus pygargus) is an important economical animal in our country which has extensive market applications. Transcriptome of roe deer antler tip was sequenced through Illumina/Solexa sequencing technology, and the transcriptome database assembly was carried out with short reads assembling program-Trinity. After transcriptome database was established, sequence alignment, functional annotation and metabolic pathway analysis with protein databases including Nr, SwissProt, KEGG and COG were done. Meanwhile. we made a simply comparative analysis with the transcriptome database of sika deer antler tip. On this basis, We choosen two functional genes in the cancer pathways named that ANXA-2 and PTN and cloned cDNA sequences including all coding regions. By making acomparative analysis with obtained the assembly cDNA sequence of these two unigenes in sequencing database of transcription group which can evidence the accuracy of the database. Carrying out the research in the level of transcriptional group is conducive to reveal the growth mechanism of roe deer antler. Therefore, this study can provide a theoretical basis to improve the yield and quality of roe deer antler.1. We generated over 50 million high quality short sequence reads by the transcriptome sequencing of roe deer antler tip. These short reads were assembled, resulting in 36865 unigenes which both ends can no longer be extended.The average length of these unigenes were 932nt, N50 was 1579nt. Nearly 98% of the sequencing quality values are in Q20 (base sequencing error rate of 1%).2. In the process of database comparison and functional gene annotation we found that the number of unigenes were 22,983 which can directly determine its CDS area and sequence direction comparing with the Nr database.The rest genes were analyzed using the software of ESTscan and results indicated that there are 510 possible proteins coding sequences. In COG functional classification 8668 unigenes were classified into 25 functional categories. In GO functional classification 18,273 unigenes were classified into 61 functional categories. Through KEGG metabolic pathway analysis 17,284 unigenes were remarked to 258 signal pathes.3. Analyzing the transcriptome of roe deer antler tip, we found collagen proteins had highest expression according to the order of FPKM value, the highest expression of the genes were COL1A1, COL1A2, COL16A1, COL9A1, COL27A1 in these collagen proteins. In addition we picked out at least 141 growth-related genes and receptors, and growth related genes of high expression are TGFB3, IGF, TGFBP, IGF4,CTGFP, PDGFR and so on. Pickde out at least 259 transcription factors, this type of high expression of genes included ATF4, TFAP1, GTFIIF, SNAI2, JunB, TFp65, etc; Picked out at least 384 species of extracellular matrix, most extracellular matrix is mainly concentrated in the collagen type composition.4. The cDNA sequences including full coding region of ANXA-2 and PTN genes were cloned, which encoded 339 and 168 amino acids. Comparing separately with sequences of the transcription group database, whose similarities are 99.7% and 99.0%, respectively.The result indicated that the transcription group sequencing result was accurate.