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Gene Cloning, Protein Expression and Functional Analysis of S100A4 of Antler Stem Cells in Sika Deer (Cervus Nippon)

Author: WangDaTao
Tutor: LiChunYi；YangFuHe
School: Jiangsu University of Science and Technology
Course: Biochemistry and Molecular Biology
Keywords: Deer antler Antler periosteum Pedicle periosteum S100A4 Retroviral Chorioallantoic membrane
CLC: S865.42
Type: Master's thesis
Year: 2010
Downloads: 40
Quote: 0
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Abstract


Deer antler as the only mammals capable of complete regeneration of the subsidiary organs , provides us with a rare biomedical model . Antler periosteum (Antlerogenic Periosteum, AP) organizations is the launching point of the deer antler development and regeneration , the corner handle periosteum (Pedicle Periosteum, PP) is a deer antler regeneration of key organizations , through the study of these two types of tissue cells to reveal antler growth regulation mechanism. S100A4 protein is a tumor-associated factors , and apoptosis , cell metastasis and angiogenesis and so on . The S100A4 protein differentially expressed between the two periosteal cells is the entry point of this article . S100A4 gene in order to study the deer antler development and regeneration process , cultivate sika deer antler periosteal cells , total RNA was extracted and anti- transcribed into cDNA Oligo dT primer . The S100A4 gene sequence (GI: 31341581) primers were designed cloned sika S100A4 gene and sequence analysis cattle . Reorganization of the the S100A4 gene sequences and expression vectors pLEGFP-C1 the recombinant vector pLEGFP-S100 and envelope vector pVSV-G transfected packaging cells GP2 -293 replication-defective virus solution containing S100A4 gene , infection pedicle periosteum cells , independent expression of the S100A4 gene introduction angle shank periosteal cells , G418 selection of infected cells to obtain stable expression of S100A4 fusion protein cell lines . RT-PCR detection S100A4 gene expression at the transcriptional level , chorioallantoic chorionic experimental analysis of S100A4 protein in deer antler development . The trial was the first cloned deer S100A4 gene and recombinant vector was constructed pLEGFP-S100 , screened pedicle periosteum cell lines stably expressing S100A4 fusion protein . Chick embryo model to detect the S100A4 protein angiogenesis close , and laid the foundation for the further reveals Velvet developmental mechanism and growth control mechanisms .

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Hunting,domestication of wild animals > A variety of wild animal domestication > Medicinal animals > Deer
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