中文摘要：

由于传统的磷酸化补齐方法对于重复序列较高的基因的合成困难较大,建立一种用于克隆全长基因的重叠延伸法并获得了成功,对全长基因进行分段扩增,从而使分段扩增片段得以重叠并互为模板,在DNA聚合酶的作用下延伸获得全长基因.根据NCB I中Ⅰ型胶原蛋白α1链结构基因第531~630个氨基酸的编码序列,设计合成了10条长度约为50 bp左右的寡聚核苷酸片段,用重叠延伸PCR法扩增合成出全长基因.PCR产物经回收后,克隆入载体pGEM-T中,并导入细菌中进行扩增.结果：经凝胶电泳和酶切鉴定、测序分析证实,合成的目的基因与设计的序列相一致.

英文摘要：

Traditional method of phosphorylation filling for the high repeat gene synthesis is quite difficult.In this paper,the establishment of a full-length gene cloning overlap extension method was successful.The full-length gene was amplified by segments,so that overlapping segments can be amplified and taken as template each other,and the full-length gene was obtained under the extension of DNA polymerase.According to the coding sequence of 531~630 amino acids of α1 type Ⅰ collagen chain gene in NCBI,ten oligonucleotide fragments at length of about 50 bp were synthesized,respectively,and the full-length gene was amplified by overlap extension PCR method.PCR products were recovered and cloned into the pGEM-T vector,and transformed into bacteria for amplification.The gel electrophoresis and restriction enzyme digestion and sequencing analysis confirmed that the synthetic gene sequence is consistent with the designed sequence.