Finer M H, Boedtker H, Doty P
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
Gene. 1987;56(1):71-8. doi: 10.1016/0378-1119(87)90159-4.
As a first step in isolating the 5' end of the chicken pro alpha 1(I) collagen gene, we constructed cDNA clones complementary to the 5' end of the pro alpha 1(I) mRNA using synthetic oligodeoxynucleotides complementary to a conserved region within the N-terminal telopeptide as primers. cDNA clones corresponding to the 5'-untranslated region, signal peptide, N-propeptide and telopeptide were identified based on homology with the human pro alpha 1(I) collagen protein sequence, and on hybridization to pro alpha 1(I) mRNA on Northern blots. A comparison of the nucleotide sequence of these clones with the sequence of the 5' end of the pro alpha 2(I) collagen mRNA confirms that there is 84% homology in a 49-bp region surrounding the translation start point, and shows that there is 70% homology in the nucleotide sequences encoding the N-propeptide triple helical region of the two type-I collagen chains.
作为分离鸡原α1（I）型胶原基因5'端的第一步，我们使用与N端端肽内保守区域互补的合成寡脱氧核苷酸作为引物，构建了与原α1（I）型mRNA 5'端互补的cDNA克隆。基于与人类原α1（I）型胶原蛋白质序列的同源性，以及在Northern印迹上与原α1（I）型mRNA的杂交，鉴定出了与5'非翻译区、信号肽、N-前肽和端肽相对应的cDNA克隆。将这些克隆的核苷酸序列与原α2（I）型胶原mRNA 5'端的序列进行比较，证实了在翻译起始点周围的49个碱基对区域内有84%的同源性，并且表明在编码两种I型胶原链N-前肽三螺旋区域的核苷酸序列中有70%的同源性。
