| 1. | The cdna encoding growth hormone ( gh ) peptide was amplified by reverse transcription polymerase chain reaction ( rt - pcr ) method using isolated total rna as template
应用逆转录-聚合酶链式反应( rt - pcr )技术克隆得到编码草鱼生长激素( cgh )的基因cdna ，并定向克隆到puc18载体上。 |
| 2. | Pcr product was cloned to the downstream of gst gene according to the right open reading frame ( orf ) in pgex - 6p - l vector , and e . coli bl21 was transformed by the recombinant plasmid for expression
Pcr产物经纯化、酶切后，按正确的阅读框架定向克隆到表达性载体pgex - 6p - 1中谷胱甘肽转移酶( gst )基因的下游。 |
| 3. | We confirmed the correct construction by pcr and restriction enzyme analysis . in this research , hypocotyls were used as the explant and several factors affecting genetic transformation of carrot mediated by agrobacterium tumefaciens were studied
利用vp7基因和质粒pbi121上相同的单克隆位点，将vp7基因定向克隆到植物表达载体pbi121上，构建了pbi121vp7表达载体。 |
| 4. | A plant expression vector was constructed by following method : s gene of 1bv and 35s promoter was cut from recornbinant pbi121 , and then the fragment was inserted into the multiclonal sites hind / bamh i in the plasmid pcambia1305 . 1
通过从重组质粒pbi121上切下s基因(连同35s启动子)片段，将该片段定向克隆到pcambia1305 . 1质粒的多克隆位点hind 、 bamh之间，构建了一种植物表达载体。 |
| 5. | For prokaryotic expression , the coding region of the cdna of the phytoene desaturase gene was cloned by pcr and inserted into a prokaryotic expression vector pet - 21a ( + ) . the gene was overexpressed in e . coli bl21 ( de3 ) and gave rise to a 63 kd mature protein in response to the iptg induction
用pcr方法扩增番红花八氢番茄红素脱氢酶( pds )编码区序列，定向克隆到表达载体pet一艺1a ( + )上，获得重组质粒pet一21a ( + ) / cspds 。 |
| 6. | In the research of transgenic fish , green fluorescent protein gene was sub - cloned to downstream of carp p - actin gene promoter that was cloned in pucusa by molecular recombination technology . thus pagfp plasmid was constructed successfully . the recombination was determined by digestion of restriction enzyme and sequencing
实验通过分子重组技术，采用定向克隆法将绿色荧光蛋白基因亚克隆到puc118a上鲤鱼-肌动蛋白基因启动子下游，构建成能在真核生物体内表达的表达载体pagfp ，经双酶酶切法序列鉴定后，回收带启动子和目的基因片段。 |
| 7. | Another 561 bp fragment of the orf of p22 gene was amplified by pcr , and digested by pst1 and xba 1 . the amplified fragment was cloned into pbudcfal to construct the recombinant plasmid pbudce4 . 1 / p22 . then transfected it into the murine macrophages by using lipofectamine , the expression of p22 - mrna in the transfected macrophages was identified by rt - pcr
同时扩增p22编码基因orf的全长561bp片段，经pst 、 xba双酶切后，定向克隆于质粒pbudce4 . 1 ，构建真核重组重组表达质粒pbudce4 . 1 / p22 ，通过脂质体介导转染入小鼠巨噬细胞raw264 . 7 ，以rt - pcr方法鉴定目的基因在巨噬细胞中的表达。 |
| 8. | Pcr products were inserted into pbv - 220 with double digestion of restriction enduonuclease . the expression vectors of pbv - a pbv - b and pbv - c were constructed by orientaional cloning . through sds - page , bioactivity and function analysis of expressed protein , the function of phaa , phab and phac was confirmed
菌株的亚克隆基因组片段中，分离出phaa 、 phab和phac三个基因片段，定向克隆至原核表达载体pbv220上，构建了三个原核生物表达载体pbv - a 、 pbv - b和pbv - c ，通过对表达载体诱导表达，表达蛋白产物的sds - page分析、生物活性与功能分析，确定了基因phaa 、 phab和phac的生物学功能。 |
| 9. | Methods : an artificial gene for fl extracellular domain cdna was synthesized by using favored genetic codons of pichia pastroris . by inserting human fl extracellular domain cdna coding 156 amino acid resi dues into pichia pastoris expression vector ppic9k containing aox1 promoter and the sequences of alpha secreting signal peptides , a recombinant expression plasmid ppic9k - fl was constructed , and integrated into the alcohol oxidase region of the host genome
为了提高外源基因的表达量，我们根据毕赤氏酵母偏爱密码子人工合成了编码fl胞外区156个氨基酸的cdna序列，目的序列被定向克隆到酵母分泌型表达载体ppic9k质粒上，构建ppic9k - fl表达质粒。 |
| 10. | The chloroplast shsp gene was screened from the cdna library of tomato flower by pcr strategy and confirmed by sequencing . but difference was found at 3 bases of the sequence from the reported in genbank . then , an integrated vector prok ii of the chloroplast shsp gene and nptii gene ( a kanamycin resistant gene ) with camv35s promoter was constructed and introduced into tomato mediated by agrobacterium tumefaciens lba4404 . transgenic tomato were screened by their ability of growing on media containing kanamycin
本实验采用pcr方法从番茄花cdna文库中克隆到叶绿体shsp基因，经测序证实与genbank中已发表的序列在编码区相差2个碱基，其中一个碱基导致1个氨基酸的改变。将叶绿体shsp基因定向克隆于带有组成性表达启动子camv35s的植物表达载体prok中，冻融法转化农杆菌lba4404 ，利用叶圆盘法对番茄进行ti质粒介导的遗传转化。 |
