| 1. | The recombinant vector was digested with tthllll and the lacz gene from e . coli was inseted in this site , the generated plasmid is designated as pltk - uni
然后定向亚克隆swha基因于多克隆位点，获得重组转移载体pltk - ha 。 |
| 2. | In our study we have cloned the osd gene from s . typhimurium by pcr , characterized the gene product and used this gene to construct asd + expression cloning vectors ptrc99a - asd
再将hpylori尿素酶b亚单位基因与尿素酶b和热休克蛋白a融合基因分别克隆入ptrc99a一asd质粒的多克隆位点之内。 |
| 3. | 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。 |
| 4. | The recombinant transfer vector pbacpak - hbmp was constructed by insertion of the hbmp coding sequences into the multiple cloning site of transfer vector pbacpak . 8 . bmn cell line was co - transfected with pbacpak - hbmp plasmid and linearized baculovirus bacpak6 dna by dosper agent
将克隆到的hbmp基因通过适当的酶切插入到转移载体质粒pbac - pak8的多克隆位点中，获得重组转移载体质粒pbacpak - hbmp 。 |
| 5. | A 1 . 5 kb apramycin resistance fragment was inserted into nru i site of aved gene and the inactivated aved gene fragment was then introduced into mcs region of phjl401 - an e . coli / streptomyces shuttle vector with conjugation function ( containing orit gene ) . as a result of above procedures , a recombinant plasmid pid03 was obtained
将1 . 5kb的安普霉素抗性基因片段插入到aved基因中的nrui酶切位点，再将此灭活的aved基因片段插入到具有接合转移功能(含有orit基因)的链霉菌?大肠杆菌穿梭质粒phjl401的多克隆位点区，由此得到重组质粒pid03 。 |
| 6. | The interest gene was inserted in the - tha l . tho 1 multiple cloning sites of donor plasimd pfastbachtb of baculovirus expression system . after analysis by restriction endonuclease and pcr , the recombinant donor plasmid gpl - fast was transforn1ed to the competent celi dhi0bac which contalns the bacmid and the helper plasmid , the recombinant bacmid gpl - bac was acquired which would express the vpl of gpv strain h l
同时将该目的基因插入到杆状病毒表达系统的供体质粒pf _ ( ast ) b _ ( ac ) htb的xba 、 xho多克隆位点间，经酶切、 pcr鉴定后，将重组的供体质粒gp1 - fast转化到含有杆状病毒和辅助质粒的dh10b _ ( ac )感受态细胞中，获得了表达gpvh1株vp1的重组杆状病毒gp1 - bac 。 |
| 7. | The gd and ge gene was subcloned into puc18 , resulting in pugdge . the fragment from pcdnas . 1 - including hcmv promoter / enhancer , mcs and neomycin resistance gene was inserted into the bamhi and bsteii restrication sites of pugdge , resulting in the universal transfer vector pgd - m - ge . the universal transfer vector pgd - m - ge has deleted the gi gene and 363bp in the 5 ' end of the ge orf of prv . there were 11 restrication sites for insertion of the foreign gene . the upstream and downstream flanking sequences were up to 1 . 25kb and 1 . 42kb . it will be useful for developing the recombinant prv expressing foreign gene ( s )
将gd 、 ge基因连接于质粒puc18获得pugdge ，缺失质粒pugdge的bamh和bste位点间391bp的片段。在此缺失位置插入来自质粒pcdna3 . 1 -的一伪狂犬病病毒gd 、 ge 、 tk基因的克隆与通用转移载体的构建段含hcmv启动子。多克隆位点和neo报告基因的片段，构建了通用转移载体ppd m pe 。 |
| 8. | Based on a 3 . 1kb pst i fragment of genomic dna of a wild s . avermitilis , a 1 . 5 kb apramycin resistance fragment was inserted into sph i site of avec gene in the 3 . 1 kb fragment , then a recombinant plasmid pc05 was obtained by introducing above inactivated avec fragment into mcs region of phjl401 . competent cells of et12567 were transformed by recombinant plasmid pid03 and pc05 respectively
以含有avec基因的3 . 1kb基因组dnapsti片段为基础，将1 . 5kb的安普霉素抗性基因片段插入到avec基因中的sphi酶切位点，再将此插入失活的avec基因片段连接到具有接合转移功能(含有orit基因)的链霉菌-大肠杆菌穿梭质粒phjl401的多克隆位点区，由此得到重组质粒pc05 。 |
| 9. | Then some necessary elements such as gfp gene , neo gene were inserted between the loxp site and the loxs 11 site and obtained a new gene targeting vector pioxgfp . on the other hand , the chicken v - ifn cdna was cloned into the vector plox between the loxp site and the lox511 site and got another gene targeting vector ploxifn
合成loxp和lox511位点，将其同方向地克隆到载体pegfp - 1的多克隆位点之内，构建成含有lox位点的通用载体? ? plox ，然后将报告基因gfp及neo基因片段克隆到载体plox上的loxp和lox511序列之间，构建成一基因打靶载体? ploxgfp 。 |
| 10. | In this study , a 1 . 7kb kpni fragment and a lacz gene expression cassette carrying the e . coli lacz gene under the control of sv40 promoter were inserted into the transfer vector pbdtk - uni ( a 277bp acci - accl fragment in the tk gene was deleted ) . the new transfer vector was called puni - lacz . the transfer vector puni - lacz and purified genomic dna of strain bartha - k61 were used to cotransfect vero cells using lipofectin transfection procedure
本研究以呈ge ~ -表型的经典弱毒疫苗bartha - k61株为亲本株，在通用prv转移载体pbdtk - uni的基础上，在其多克隆位点中插入由sv40早期启动子控制下的lacz基因表达盒，同时将下游同源臂增加了一个1 . 7kb的kpni片段，使上下游同源臂的长度都超过了1kb ，构建了一个新的转移载体puni - lacz 。 |
