中国人β神经生长因子前体基因的克隆及其序列分析

CLONING AND SEQUENCING OF THE GENE FOR PREMATURE β NERVE GROWTH FACTOR

从正常人外周血白细胞中提取基因组ＤＮＡ，用ＰＣＲ扩增神经生长因子（ＮＧＦ）β亚基前体的全长编码区序列，将其克隆到Ｔ－ｖｅｃｔｏｒ（原始质粒为ｐＢｌｕｅｓｃｒｉｐｔⅡＳＫ（＋））上，取两个独立的克隆采用自动测序仪进行双链ＤＮＡ双向测序，结果表明：两个克隆的序列完全相同，该序列与国外报道的ＮＧＦ序列有一个碱基的差别，从而导致ＮＧＦ前导肽中一个氨基酸的改变，而成熟的ＮＧＦ序列没有改变。

The entire coding sequence of premature nerve growth factor (NGF) β subunit was amplified from Genomic DNA extracted from peripheral white blood cells of Chinese by polymerase chain reaction (PCR). The PCR products were cloned into a premade T vector (linearized plasmid pBluescript Ⅱ SK(+) with a T base at 3′ terminal), followed by automatic sequencing with double strand plasmids. The nucleotide sequence coding for the mature NGF is the same as that reported by Europeans, but there is a base difference within the proper region, resulting in an amino acid replacement in premature NGF.