Lovell-Badge R H, Bygrave A, Bradley A, Robertson E, Tilly R, Cheah K S
Proc Natl Acad Sci U S A. 1987 May;84(9):2803-7. doi: 10.1073/pnas.84.9.2803.
Type II collagen is crucial to the development of form in vertebrates as it is the major protein of cartilage. To study the factors regulating its expression we introduced a cosmid containing the human type II collagen gene, including 4.5 kilobases of 5' and 2.2 kilobases of 3' flanking DNA, into embryonic stem cells in vitro. The transformed cells contribute to all tissues in chimeric mice allowing the expression of the exogenous gene to be studied in vivo. Human type II collagen mRNA is restricted to tissues showing transcription from the endogenous gene and human type II collagen is found in extracellular matrix surrounding chondrocytes in cartilage. The results indicate that the cis-acting requirements for correct temporal and spatial regulation of the gene are contained within the introduced DNA.
II型胶原蛋白对脊椎动物形态的发育至关重要，因为它是软骨的主要蛋白质。为了研究调节其表达的因素，我们在体外将一个包含人类II型胶原蛋白基因的黏粒导入胚胎干细胞，该基因包括4.5千碱基的5'侧翼DNA和2.2千碱基的3'侧翼DNA。转化后的细胞对嵌合小鼠的所有组织都有贡献，从而可以在体内研究外源基因的表达。人类II型胶原蛋白mRNA局限于显示内源性基因转录的组织，并且在软骨中软骨细胞周围的细胞外基质中发现了人类II型胶原蛋白。结果表明，该基因正确的时间和空间调节所需的顺式作用元件包含在导入的DNA中。
