This method has several strong points : ( 1 ) eliminating the possibility of ringing self of vector . ( 2 ) the inserting fragments ca n ' t ligate each other . ( 3 ) the translating rate with the partially filled in method is equal to phosphatase method 末端半补齐技术的优点有： ( 1 )彻底消除载体自环化的可能性； ( 2 )消除了插入片段自身相互连接的可能性； ( 3 )同常用的碱性磷酸酯酶法相比较，采用半补齐技术其连接产物的转化率不受影响。
The construction of pci - il - 2 : primers were designed by the software oligo , xhol and sail were added to the primers , the cdna of il - 2 was obtained by pcr . the cdna of il - 2 was ligated with pci - neo cleaved by xhol and sail . the recombinant was evaluated by pcr Pcr法扩增几亿片段，在t4连接酶的作用下与xhol和sal工酶切的pci neo载体连接， pcr法鉴定重组体，用xho工， xhol sal ， ban工工酶切进一步验证重组体，命名为pclll 2 。
A 12 bp sequence of the 5 " end from the polyhedrin protein gene of bmnpv was ligated to the 5 ' end of hbsag ( pres2 + s ) protein coding sequence by pcr . the fusion product coding for hbv surface antigen medium sized ( hbmp ) with 4 - additional aa of bmnpv polyhedrin protein was obtained 本研究通过pcr突变的方法，在hbsag ( pres2 + s )前s2序列的5 ’端融合了bmnpv多角体蛋白基因5 ’端的12个碱基，获得了融合乙肝表面抗原中蛋白基因( hbmp ) 。
The constuction of plant expression vector , tissue culture of cabbage and gene transformation conditions were studied to obtain ubic transformants . ubic gene was obtained from the escherichia coli . by pcr amplication and was ligated to puc118 vector 为获得转ubic基因甘蓝，对植物基因工程载体的构建、甘蓝的组织培养以及基因转化条件进行了研究。通过pcr方法从大肠杆菌基因组中扩增得到了ubic基因，扩增产物克隆到puc118载体，转化大肠杆菌jm109 。
Conclusion : by restriction enzyme secting , ligating , transforming , restriction enzyme analysis , and final dna sequencing , the pbd - i and pbd - ii gene were proved to be recombinated with the expression vector and the recombinated vector ppd - 1 and ppd2 were transformed successfully 结论：经过酶切、连结，构建成重组质粒ppd上、 ppd上，再经转化、抽提质粒及酶切分析，最后经dna测序证实， rticr扩增的pbd i 、 pbd 11基因与piflpdt ” xsi表达载体构建成功。
Ts87 gene fragment was picked out by screening adult t . s cdna library using sera of rabbit raised against soluble antigen of t . s and using sera of rabbit infected artificially with t . s . the ts87 fragment was ligated to vector pcdnas . l , which contains a cmv promoter / enhancer 本研究采用通过免疫筛库得到的旋毛虫抗原基因片段ts87与载体pcdna3 . 1连接。该载体含有人类巨细胞病毒( cmv )的高效启动子和增强子，是一种外源基因在哺乳动物细胞内高效表达的理想载体。
The study on the function and mechanism of phrip1 is important for clarifying how the cell plate and cell wall form in plants . in this study , full length of phrip1 is amplified by pcr and ligated into pks plasmid , then the bait plasmid , peg202 - phrip1 , is constructed . the inseret gene are sure to be translated into the right fusion protein through its sequence . in the yeast two - hybrid system , the bait plasmid ( peg202 - phrip1 ) and a reporter plasmid ( psh18 - 34 ) are introduced into the yeast ( egy48 ) by co - transformation . then cdna library ( which is in pjg4 - 5 ) is screened and two genes are obtained . the two insert gene fragments are sequenced . one of them is plastocyanin , the other is putative photosystem i reaction center subunit ii precursor , both of them are the necessary components of photosynthetic chain 成膜素相关蛋白1 ( phrip1 )是一个含608个氨基酸的蛋白质，它对于植物胞质分裂中细胞板的形成起到了十分重要的作用。研究phrip1的功能和机制，对在分子水平上阐明植物细胞板以及细胞壁形成的机理具有重大的生物学意义。在本实验中，根据phrip1的序列设计引物对其进行pcr扩增，得到该基因后将其连接到了pks质粒上，并进一步构建成了诱饵质粒peg202 - phrip1 。
In order to get the soluble recombinant eo protein and inspect the protein expression status convinently , the egfp and eo gene were ligated into baculovirus transfer vector . with the co - transfecting sf9 cells of baculovirus recombinant transfer vector and linearized viral dna , and plaque purification in the posttransfection procedure , the pure recombinant baculovirus were harvested , which infected the sf9 cells for amplifying to generate a p - l stock . . in the meantime , the fluorescence microscopy detection indicated expressed egfp protein to confirm the heterogenous protein expression of recombinant baculovirus . the pi - stock from a pure plaque was used to generate a high liter p - 2 stock , which was determined in liter as 1 . 14 107pfu / ml by performing a plaque assay . when a volume of p - 2 stock infected the sf9 cells with moi 5 - 10 for expression , the strong fluorescence was obeserved on the day 3 of postinfection 此外，为了得到可溶性重组eo蛋白并便于观察重组蛋白的表达情况，我们将egfp基因与eo基因相连插入昆虫杆状病毒转移载体中，与线性杆状病毒dna共转染sf9细胞后通过噬斑纯化得到纯的重组杆状病毒，将其感染sf9细胞制备p1种子液，同时用荧光显微镜观察绿色荧光蛋白的表达情况剔除表达效果差的重组杆状病毒。再用p1种子液感染sf9细胞制备高效价的p2种子液。通过病毒液的梯度稀释和噬斑测定，确定p2种子液的病毒滴度达1 . 14 10 ~ 7pfu ml 。
The library was rescued with phage m13k07 in order to display scfv on the surface of the phage and to form the recombinant phage antibody library . one of positive scfv clones , named pcsal , was selected with phage - elisa after panning and screening by bull sperm three times . scfv fragment , amplified from pcsa1 , was ligated to pmd18 - t vector for sequencing analysis 取阳性重组噬菌体抗体克隆株pcsa1 ， pcr扩增其scfv基因，筛选重组子进行序列测定，发现其序列符合小鼠抗体基因的一般特征，并且与几株抗磷酸胆碱的抗体重链和轻链可变区序列的同源性达80以上；推测pcsa1scfv针对的抗原是磷酸胆碱类物质。
