1.RFLP analysis with single restriction enzyme of PstⅠ, HindⅢ, MluⅠ, or BglⅡand double enzymes of HindⅢand MluⅠwere performed with Southern blot of digoxigenin-labeled probes targeting to gene zot, ace, cep, respectively. Results Two close ribotypes were found in the 6 strains, whose 5 strains had nct-CTXΦgenomes, but neither CTXΦnor nct-CTXΦgenome was found in another strain.
以zot、ace、cep基因探针South- ern blot分别检测nct-CTXΦ样基因的PstⅠ、HindⅢ、MluⅠ、BglⅡ酶切及HindⅢ、MluⅠ双酶切的限制性酶切长度多态性,结合拷贝间相邻片段HindⅢ酶切位点分析,构建nct-CTXΦ样基因组物理图谱。
2.Beam walking test (BWT) measuring the latency of different beam was used to detect the changes of coordination and balance function; Footprint pattern test (FPT) was used to detect the changes of gait, including left/right stride length (LSL,RSL), front-base width (FBW), hind-base width (HBW), distance from left/right front footprint/hind footprint overlap (DLFHFO,DRFHFO);
应用Footprint pattern test(FPT)记录左/右侧步长(left/right stride length,LSL,RSL),前肢间距离(from-base width,FBW),后肢间距离(hind-base width,HBW)及左/右侧前、后爪心间距离,包括(distance from left/right from footprint/hind footprint overlap,DLFHFO,DRFHFO)检测各组大鼠步态的变化;
3.The monomer ( about 350 bp )of the Hind Ⅲ satellite DNA was isolated from the genomic DNA, and named as CDa(HindⅢ)-1. The Southern Blot experiments with this fragment as a probe indicates that the Hind Ⅲ Satellite DNA consists of tandemly repeated monomers.
以α-~(32)P标记的这种片段与经HindⅢ部分消化的基因组DNA进行Southern印迹杂交,杂交图谱显示出典型的从单体到多聚体分布的“梯状”区带,证明CDa(HandⅢ)-1片段是前后串接式重复的卫星DNA.
4.Methods The open reading frame of HBV X gene from HBV genome (subtype adw) was recovered from pEcob6 that was digested with restricted enzyme and subcloned into pBluscript KS?+. The resulting plasmid pBKS?X was digested with Hind Ⅲand NotⅠ, and the X fragment was ligated with the large fragment of pCEP4, which was also digested with Hind Ⅲ and NotⅠ.
方法 用限制性内切酶从p Ecob6 上切下 X 基因读码框，克隆到p B K S＋ 上，再将 X 片段插入p C E P4 的 Hind Ⅲ和 NotⅠ位点间。
5.2 Synthesized completed p27 cDNA by PCR, and cloned it into pUC19 for DNA sequence analysis, pUC19-p27 and pcDNAS were digested with Hind III and EcoR I, then the p27 cDNA was lignated with pcDNAS at Hind III and EcoR I restriction sites by T4 lignase, and the eukaryotic expression plasmid pcDNAS-p27 for tumor suppression gene p27 formed;
2．．PCR法从含有p27全长的质粒中合成p27 cDNA片段，连接酶连接至pUC19质粒进行测序，证明PCR产物序列完全正确，限制性内切酶HindⅢ与EcoRI酶切pUC19-P27,获得p27 cDNA片段，连接酶连接至pcDNA3，克隆出真核表达质粒；
6.Restriction enzyme mapping of AFPgene in genomic DNA with EcoRI,BamHI. HindⅢ and PstI,followed by Southernblotting and hybridization with ~(32)P-RAF 65and ~(32)P-RAF87 probes,showed rather dif-ferent banding pattern in Hindlll and PstIdigestion but no difference in EcoRI andBamHI. This indicates some gross structuralchanges of AFP gene in AH_(66).
以BamHI,EcoRI,HindⅢ和PstI酶解基因组DNA,然后与缺口翻译标记的~(32)P-RAF_(65)和~(32)P-RAF_(87)探针杂交,测知BamHI和EcoRI的酶谱相同,而HindⅢ和Pst I的带型明显不同,表明结构上存在某些变化。
7.METHODS: The whole expressive fragment of hTM gene was amplified by PCR from human genome. Both hTM gene and pcDNA3.1(+)/neo empty vector was digested by HindⅢ and EcoRⅠ. Two digested fragments were ligated into pcDNA3.1/hTM with T_4DNA ligase.
方法 :PCR法扩增hTM基因的全长表达片段 ,HindⅢ /EcoRⅠ酶切后与pcDNA3 1(+) /neo质粒连接成pcDNA3 1/hTM。
8.Wuchang Fish (M. amblycephala) liver mtDNA was cleaved by the restriction endonuclease. Upon complete digestion numbers of restriction fragments of BamHI, BglⅡ, BglⅠ, EcorⅠ, HindⅢ and HpaⅡ were determined to be 2,2,3,3,3 and 7 respectively.
武昌鱼肝线粒体(mt)DNA经六种限制性内切酶BamHI,BgⅢ,BgⅡ,EcoRI,HindⅡHpall单酶完全酶解分?e得到2,2,3,3,3和7个片段。
9.Recombined plasmid after separation with Sepharose 2B chromatography was cut with restriction endonuclease E CORI and Hind Ⅲ . Results of analysis with electrophresis showed the purity of plasmid separated with Sepharose 2Bchromatography could fulfill the analysis of restriction map for clone fragments .
本文对Sepharose 2B层析分离后的重组质粒pGEM-1用E CORI及HindⅢ限制性内切酶进行切割,电泳分析结果表明用Sepharose 2B层析分离的质粒纯度可满足于克隆片段的酶切图谱分析。
10.The recombinant plasmid pUC18/TK containing the full length cDNA coding herpes simplex virus type I(HSV 1) thymidine kinase was digested by the EcoR I and the Hind III restriction endonucleases.
用EcoR Ⅰ和Hind Ⅲ双酶切已构建的含单纯疱疹病毒I 型胸苷激酶（ HSV?1 TK） 基因的p UC18／TK 质粒，将切出的TK 基因片段克隆入原核表达载体p WR450?1 中，构建成HSV?1 TK 基因重组表达质粒p WR450?1／TK。
