权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Isolation of novel candidate genes involved in bovine adipocyte differentiation and marbling using a clonal bovine intramuscular preadinocyte (BIP) cell line

使用克隆牛肌内前脂肪细胞 (BIP) 细胞系分离参与牛脂肪细胞分化和大理石花纹的新候选基因

基本信息

批准号：
14360162
负责人：
ASO Hisashi
金额：
$ 10.37万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

To order to isolate candidate genes involved in bovine adipocyte differentiation, we have constructed a subtraction library from a clonal bovine intramuscular preadipocyte (BIP) cell line using the suppression subtractive hybridization method. We have isolated a set of subtracted cDNA fragments whose respective mRNA levels are up regulated during the adipogenic differentiation of BIP cells, and cloned a full-length cDNA from differentiated-BIP-lambda ZAP II cDNA library. The clone 4-3 was isolated and was highly homologous to the sequence of human, mouse and rat collagen prolyl 4 hydroxylase alpha III subunit (C-P4HIII). C-P4HIII is known to refold collagens and collagen like proteins, and as necessary for folding collagen triple helices structure. From the expression analysis of C-P4HIII mRNA and protein during adipogenesis in BIP cells, C-P4HIII expression was elevated both mRNA and protein. It is known that extracellular matrix (ECM) of cell surface has been restructured during adipogenesis. The ECM was decomposed by adipogenic induction, and up-regulated expression of components, included collagens for the adipocyte. These results suggest that C-P4HIII may be associated with adipocyte differentiation and adipose formation in cattle and is a potentially useful marker for adipogenesis.

为了分离与牛脂肪细胞分化相关的候选基因，我们利用抑制减法杂交的方法从克隆牛肌内前脂肪细胞（BIP）细胞系中构建了减法文库。我们分离了一组在BIP细胞成脂分化过程中各自mRNA水平上调的缺失cDNA片段，并从分化的BIP-lambda ZAP II cDNA文库中克隆了一个全长cDNA。克隆4-3与人、小鼠和大鼠胶原脯氨酸4羟化酶α III亚基（C-P4HIII）序列高度同源。已知C-P4HIII可以折叠胶原蛋白和胶原样蛋白，并且是折叠胶原蛋白三螺旋结构所必需的。通过对BIP细胞脂肪形成过程中C-P4HIII mRNA和蛋白的表达分析，C-P4HIII mRNA和蛋白的表达均升高。在脂肪形成过程中，细胞表面的细胞外基质（ECM）发生了重构。ECM被诱导成脂分解，脂肪细胞中胶原蛋白等成分表达上调。这些结果表明，C-P4HIII可能与牛的脂肪细胞分化和脂肪形成有关，是脂肪形成的潜在有用标记物。