中文摘要：

目的 体外拼装带c-myc分子标签的人三叶因子家族2（human trefoil factor family2，hTFF2）的融合基因c—myc—htff2，并构建其克隆载体pBS-TFF2，以便为进一步构建TFF2各种表达载体打下基础。方法根据hTFF2氨基酸序列，结合乳酸链球菌氨基酸表达密码子，设计编码htff2的基因序列（cDNA）；在其上游添加c-myc碱基序列作为分子标签，设计融合基因c—myc—htff2；参照pBluescript Ⅱ sk（＋）载体的酶切位点碱基序列，分别在c-myc—htff2的5’和3’端设计Sal Ⅰ和Bam H I酶切位点；用DNAWORKS在线程序指导，将c-myc—htff2设计成14个相互互补的寡核苷酸片段，经PCR扩增得到目的基因片段c—myc—htff2；再将目的基因片段亚克隆至pBluescript Ⅱ sk（＋）载体，进行酶切鉴定及DNA测序。结果 成功地在体外拼装了c—mye-htff2基因，完成了pBS-TFF2构建，且DNA测序结果与设计序列完全一致。结论 在DNAWORKS程序指导下设计寡核苷酸并进行体外基因拼装是合成目的基因的有效方法；成功构建了c-myc-htff2融合基因的克隆载体pBS-TFF2。

英文摘要：

Objective To assemble fusion gene of c-myc tagged human trefoil factor family 2 （hTFF2） in vitro and construct a cloning vector of this fusion gene. Methods Based on amino acid sequence of hTFF2 and codon of lactococcus lactis, cDNA of htff2 sequence was designed and extended at their 5＂ ends with a sequence encoding the c-myc tag and the sequence of fusion gene c-myc-htff2 was designed. According to restriction enzyme sites of pBluescript Ⅱ sk （＋）, the SalⅠ and BamH Ⅰ were arranged at 5＂ and 3＂ ends of the fusion gene respectively. Instructed by DNAWORKS program, the fusion gene sequence of c-myc-htff2 was designed as 14 oligonucleotides that overlapped each other. The target gene fragment of c-myc-htff2 fusion gene was obtained by the means of polymerase chain reaction （PCR） based gene assembly. Then, the c-myc-htff2 was subcloned into vector of pBluescript Ⅱ sk （＋） for identification of the fusion gene by restricting enzyme excision and DNA sequencing. Results Assembly of c-myc-TFF2 fusion gene in vitro and construction of pBS-TFF2 had been completed successfully and DNA sequencing showed that the sequence of synthetic gene accorded with the expectation. Conclusion With the help of DNAWORKS program, the design and assembly of oligonucleotides in vitro is effective to synthesize a target gene, and the cloning vector of c-myc-htff2 fusion gene has been successfully constructed.