大熊猫线粒体DNA的九种限制酶图谱

本文用9种限制性内切酶（BamHⅠ,BglⅠ,BglⅡ,EcoRⅠ,EcoRⅤ,PstⅠ,PvuⅡ,SalⅠ,XhoⅠ）分析大熊猫的线粒体DNA（mtDNA）。构建其中5种酶（BamHⅠ,EcoRⅠ,EcoRⅤ,PstⅠ,PvuⅡ）的mtDNA物理图谱。大熊猫mtDNA的分子大小约为16.4 Kb,酶切位点是随机分布。我们的结果为进一步研究大熊猫mtDNA进化提供了基础资料。     

鲤鱼肝组织线粒体DNA的限制性内切酶分析

用EcoRⅠI,HindⅢ,PstⅠ,BglⅡ,BamHⅠ,Xho Ⅰ, Xba Ⅰ, Sal Ⅰ和Kpn Ⅰ共9种限制性内切酶酶切对鲤鱼肝组织的mtDNA进行酶解,利用Gel-Pro Analyzer算出各酶切片段长度及mtDNA大小,测出其大小约为16．61kb(1kb=1×103b),另外,对电泳条件的优化进行了探讨,并将实骚结果与以前报道的有关酶切实验结果进行了比较,表明鲤鱼mtDNA的长度及限制性酶切位点均存在地域差异．     

鳜及大眼鳜线粒体DNA比较研究

采用10种限制性内切酶HindⅢ、EcoRⅠ、BglⅠ、BglⅡ、BamHⅠ、PvuⅡ、PstⅠ、SalⅠ、XbaⅠ、XhoⅠ对鳜及大眼鳜肝脏线粒体DNA（mtDNA）进行了分析。鳜鱼mtDNA分子量为9.703×106u,大小为16．19kb;大眼鳜mtDNA分子量为9．603×106u,大小为16．02kb。根据单、双酶切结果构建了鳜及大眼鳜mtDNA10种酶限制性酶切图谱,并对两种鱼的mtDNA酶切图谱进行了比较。     

四个鲫鱼品系线粒体DNA的限制性酶切分析

用差速离心和核酸酶消化法从红鲫 (C auratusredvar .)、湘鲫 [F1hybridsofredcruciancarp (♀ )×commoncarp (♂ ) ]、野鲫 (C auratusauratus)和白鲫 (C auratuscuvieri)的肝组织及白鲫的卵巢中提取和纯化线粒体DNA。用 9种内切酶 (EcoRⅠ、HindⅢ、PstⅠ、BglⅡ、BamHⅠ、XhoⅠ、XbaⅠ、SalⅠ和KpnⅠ )进行单酶酶解 ,经琼脂糖凝胶电泳分析 ,检测出PstⅠ、KpnⅠ和BglⅡ 3种酶在品系间存在限制性片段长度多态性 ,但并未检测出品系内的限制性片段长度多态性。计算出红鲫、湘鲫、白鲫和野鲫的mtDNA大小分别约为 16 19、 16 0 2、 16 6 0和 16 0 6kb。根据限制性酶切片段共享度 ,计算出 4个品系间的遗传距离 ,结果表明存在直接亲缘关系的红鲫与湘鲫之间的遗传差异最小 ,证实了红鲫与子代湘鲫之间mtDNA遵循母系遗传的特性。     

鸡肝脏线粒体DNA的限制图谱

本文用六种限制性内切酶对鸡肝脏线粒体DNA(mtDNA)进行了酶解。Eco RⅠ、Bam HⅠ、SalⅠ、HindⅢ、BglⅠ在鸡肝mtDNA上分别有2、2、3、4、4个切点,BglⅡ不能切割鸡肝mtDNA。根据鸡肝mtDNA的单酶、双酶完全酶解以及部分酶解片段的分子量,建立了鸡肝mtDNA的限制图谱。     

树qumtDNA限制性内切酶图谱及其与小家鼠褐家鼠的新缘关系

本实验用ApaⅠ,AvaⅠ,BamHⅠ,BclⅠ,BglⅠ,ClaⅠ,EcoRⅠ,EcoRⅤ,HpaⅠ,PstⅠ,PvuⅡ,ScaⅠ,XbaⅠ等13种限制性内切酶分析树qu(Chiromyscus chiropus) 的mtDNA限制性片段长度多态性,并用双酶解法构建了其中8种酶的限制性内切酶图谱。根据限制性片段差异法和分子钟,计算并讨论树qu和小家鼠(Mus musculus)、褐家鼠(Rattus norvegicus)的mtDNA遗传距离和亲缘关系。结果表明树qu与褐家鼠的关系较接近,两者的分歧时间在距今1500-2000万年前,即处于中新世早中期。     

条纹斑竹鲨线粒体DNA的研究

用6种限制性内切酶分析了4条条纹斑竹鲨的线粒体DNA(mtDNA)。PstⅠ、Hpa Ⅰ、XbaⅠ、EcoRⅠ、EcoRⅤ、BglⅡ在条纹斑竹鲨mtDNA分子上分别具有0至2个切点, mtDNA分子大小为16.6kb,根据单酶和双酶完全酶解片段的大小,构建了条纹斑竹鲨mtDNA 的限制性酶切图谱。 Abstract:Mitochondrial DNA(mtDNA)form 4 samples of Chiloscyllium plagiosum was analyzed by 6 kinds of restriction.The number of cleavage sites were as follow:2 for HpaI,XbaI and EcoRI respectively;1 for BglII and EcoRV respectively;None for PstI.Molecular size of mtDNA was found to be 16.6kb.According to analysis of single and double enzyme cleavage,the map of restriction enzyme was constructed.     

创新霉素产生菌中质粒的研究——Ⅳ.AJP1质粒的物理图谱

用MAK法提纯创新霉素产生菌——济南游动放线菌中天然质粒AJP1,并对其进行了酶解分析。实验表明,限制性内切酶BamHⅠ、BglⅡ、HindⅢ、KpnⅠ、SstⅠ和HpaⅠ不能降解AJP1,XhoⅠ、PsiⅠ、XbaⅠ、EcoRⅠ、PvuⅠ、SinaⅠ和SalⅠ在AJP1上分别有1、1、1、2、3、5,和7个切点。应用双酶解等方法进行酶切位点的定位,绘制了共有20个酶切位点的AJP1物理图谱。     

疫苗

960999 禽痘病毒DNA复制及作图分析[英]/Zantinge, J. L.…//Can. J. Microbiol. -1995,41(4～5).-378～387[译自DBA,1995,14(17),95-10127] 产生了一种禽痘病毒Chick-N-Pox疫苗株DNA基因组的EcoRⅠ、BamHⅠ、NcoⅠ和PstⅠ限制性物理图谱。利用琼脂糖凝胶电泳和PAGE分析了EcoRⅠ、BamHⅠ、BglⅡ、NcoⅠ、StyⅠ、PstⅠ和PvuⅡ消化DNA的凝胶分布结果。从中估测禽痘病毒基因组的     

金秋梨和新高梨的分子遗传学对比分析

金秋梨和新高梨的形态性状、生物学性状以及酯酶同工酶、过氧化物酶同工酶、细胞色素氧化酶同工酶酶谱均产生了变化。利用异源DNA探针—小麦rDNA克隆pTA71,与四种核酸内切酶PstⅠ、BamHⅠ、EcoRⅠ及BglⅡ酶切消化的金秋梨和新高梨叶片总DNA杂交,结果表明,pTA71-PstⅠ和pTA71-BamHⅠ二种探针—酶组合可在金秋梨和新高梨中检测到RFLP差异。     

鲤鱼mtDNA酶切图谱的构建

采用密度梯度离心法及ＲＮａｓｅ消化法制备并纯化了鲤（ＧｙｐｒｉｎｕｓｃａｒｐｉｏＬｉｎｎａｅｕｓ）肝脏线粒体ＤＮＡ（ｍｔＤＮＡ）,用１０种限制性内切酶对ｍｔＤＮＡ进行了分析,鲤鱼ｍｔＤＮＡ分子量约１０．１２×１０￣６,约１６．４９ｋｂ．ＳａｌⅠ、ＰｓｔⅠ、ＢａｍＨⅠ、ＸｂａⅠ、ＢｇｌⅠ、ＰｖｕⅡ、ＸｈｏⅠ、ＥｃｏＲⅠ、ＤｒａⅠ和ＨｉｎｄⅢ分别为１、１、３、３、３、４、１、４、４、和６个切点。根据单酶解及双酶解结果,构建了鲤ｍｔＤＮＡ１０种具酶３０个切点的限制性酶切图谱。     

人乙型肝炎病毒(HBV)adr亚型基因组的多态性

由乙型肝炎adr亚型病毒(HBVadr)携带者26人的混合血清,得到了HBVadr基因组克隆株(PADR)158株,对这些克隆株进行四种限制性内切酶(BglⅡ,HindⅢ,PstⅠ,XhoⅠ)切点测定,并对其中S株的13种限制性内切酶图谱进行比较研究,发现同为adr亚型病毒,其基因组的限制性酶切图谱存在差异。另外,通过HindⅢ)切点得到的12个克隆株(PADR-H),也进行了酶切图谱分析。在这170个克隆株中,已经发现了5种类型的HBVadr基因组限制性酶切图谱,其中有6种酶(AvaⅠ,EglⅠ,BglⅡ,HincⅡ,HindⅢ,HpaⅠ)的7个变异点。本文报道了HBVadr基因组的多态性现象。     

Restriction endonuclease cleavage map of mitochondrial DNA from Aspergillus nidulans.

Mitochondrial DNA of the ascomycete fungus Aspergillus nidulans, a circular molecule of 31 500 base pairs, is cleaved by restriction endonucleases Eco R I, Hind II, Hind III and Bgl II into 3, 7, 9 and 5 fragments, respectively. The relative positions of the cleavage sites could be mapped by analysis of fragments obtained by double enzyme digestions of whole DNA and by complete and partial redigestion of isolated restriction fragments.     

Digestion-after-shuffling: a method for combinatorial library construction

We have developed a convenient method for family shuffling of amino acid sequences, termed digestion-after-shuffling. After DNA shuffling of homologous genes, plasmid mixture is extracted from a library and used for several double digestions with restriction enzymes. For each double digestion, two restriction enzymes are selected, corresponding to the single restriction sites of different parental genes. After digestions, fragments with expected sizes are obtained by gel purification and religated to construct recombinant plasmids. Thus, the obtained genes should be chimeras and have at least two restriction sites originating from different parental sequences.     

The cDNA Cloning and Nucleotide Sequence of the Gene Encoding Nonstrucure Protein of Rice Dwarf Virus Genome Segment 10

The results of cloning and sequencing the gene encoding nonstructure protein of the rice dwarf virus (RDV) gtnome segment 10 with polymerase chain reaction(PCR) technique were reported. The amplified PGR product was cloned into Hind Ⅱ site of plasmid pGEM3zf(-) and analysed with restriction enzymes. The physical map of the cloned fragment has been constructed, the insert is 1150 bp in length with restriction enzyme sites of Sac Ⅰ, Hind Ⅲ, NdeⅠ, BamH Ⅰ, etc. Besides, two restriction enzyme sites Bgl Ⅱ and EcoR Ⅰ have been separetely added in the 5 and 3 end of the segment in order to be cloned into plant intermediate vector in a convenient way. The fragments cleaved by the above-mentioned restriction enzymes were subcloned and the DNA sequence of full length of segment 10 was determined. In comparison with the RDV epidemic in Japan, the nucleotide sequence and deduced amino acid sequence of cloned segment 10 are 96.03% and 97.17% in homology respectively.     

Restriction endonuclease map of Euglena gracilis chloroplast DNA.

A physical map of the Euglena gracilis chloroplast genome has been constructed, based on cleavage sites of Euglena gracilis chloroplast DNA treated with bacterial restriction endonucleases. Covalently close, circular chloroplast DNA is cleaved by restriction endonuclease SalI into three fragments and by restriction endonuclease BamHI into six fragments. These nine cleavage sites have been ordered by fragment molecular weight analysis, double digestions, partial digestions, and by digestion studies of isolated DNA fragments. A fragment pattern of the products of EcoRI restriction endonuclease digestion of Euglena chloroplast DNA is also described. One of these fragments has been located on the cleavage site map.     

Physical mapping of plasmid pDB101: A potential vector plasmid for molecular cloning in streptococci

A physical map of the streptococcal macrolides, lincomycin, and streptogramin B (MLS) resistance plasmid pDB101 was constructed using six different restriction endonucleases. Ten recognition sites were found for HindIII, seven for HindII, eight for HaeII, and one each for EcoRI, HpaII, and KpnI. The localization of the restriction cleavage sites was determined by double and triple digestions of the plasmid DNA or sequential digestions of partial cleavage products and isolated restriction fragments, and all sites were aligned with a single EcoRI reference site. Plasmid pDB101 meets all requirements essential for a potential molecular cloning vehicle in streptococci; i.e., single restriction sites, a MLS selection marker, and a multiple plasmid copy number. The vector plasmid described here makes it possible to clone selectively any fragment of DNA cleaved with EcoRI, HpaII, or KpnI, or since the sites are close to each other in map position, any combination of two of these restriction enzymes.     

玉米黑粉菌mtDNA的研究 Ⅱ.限制性内切酶酶切图谱

本文报道玉米黑粉菌mtDNA的限制性内切酶酶切图谱。分别将mtDNA的Bam HI各片段制成探针,与mtDNA分别用8种酶酶切后的Southern膜进行杂交,用片段重叠法得出各套片段的排列次序,再将克隆化的Bam HI片段进行第二酶切,按分子量拼排出各酶酶切位点在mtDNA上分布的图谱。此外,片段重叠分析时,还发现玉米黑粉菌mtDNA为环状结构;杂交分析时还发现mtDNA内没有明显的重复序列。DNA总长60.7kb。     

A physical map of the genome of Ureaplasma urealyticum 960T with ribosomal RNA loci.

A physical map is presented for the 900 kilobase pair genome of Ureaplasma urealyticum 960T, locating 29 sites for 6 restriction endonucleases. The large restriction fragments were separated and sized by pulsed-field agarose gel electrophoresis (PFGE). Their locations on the map were determined by probing Southern blots of digests with individual fragments isolated from other digests and by correlating the products of double digestions and partial digestions. An end-labelling technique was used to detect small fragments not readily observed by PFGE. Two loci for rRNA genes have been determined by probing with cloned DNA.