PCR-RFLP基因分型方法(1).pptxVIP

PCR-RFLP 基因分型方法 关于序列查找、引物设计和酶切位点分析 限制性片段长度多态性(restrition fragment length polymophism,RFLP): 是指由限制性酶切位点间的插入、缺失、重排或 点突变所引起的基因型间限制性片段长度的变异。 PCR-RFLP: 是在PCR 和 DNA 序列分析基础上产生的 RFLP 技 术。该方法是通过PCR 扩增一段 DNA 片段，然后再 选择适当的限制性内切酶，消化PCR 产物，经电泳， 可得到有特异性的电泳谱带，从而达到鉴定不同基因型 的目的。 聚合酶链式反应连接的限制性片段长度多态性分析 琼脂糖电泳 H: 突变杂合子 N: 野生型 M: 突变纯合子 野生型含限制性酶切位点 突变型限制性酶切位点消失 H N M 限制性核酸内切酶酶切 Step by step 基 因 型 电 泳 选择内切酶 设计引物 确定基因序列 Swiss-Prot Protein knowledgebase TrEMBL Computer-annotated supplement to Swiss-Prot /sprot/ 基因序列查找 /Genbank http://www.genomatix.de/ NCBIGenBank Overview Bt Toxin Noinenlature Introdrd:an Racent Nwg updated 28/8m9 Yio Hoeclate updated 16/109 Holotype Tgang updated 9ang Eul Taain List updated 23/8/09 Threa Domun Dendrotn(pd) updated 28/11/08 (al进tonris·prints on legal sixe paper) Thees Doman Dendroaum(pdii updated 21 (eneluding tettiary ranks-prnte on A4 Qthet Toang Dendroesnpt updated 13/12/01 Commtttt Bacillus thuringiensis Toxin Nomenclature: The Baciflus tharingiensis dells-endotomn nomenclature commdttee was set up in 1993 am order to update the no Hofte and Whiteley (Microbiological Reviews 53.242-255)and consists of the following sg The curent nomenclature,based solely on amino acid identity,allows closely related toains to be ranked together to bioassay each new toxin againat a growing seriea of organisms.Biological epeciffeity being a component of th have also been erchanged for Arabie numerals in the pnmary rank(eg CrylllA became Cry3A).Each new torin is a tanks.A completoly new toxin might therefore be assigned the name Cry50Aal.For the sake of convenience we n (which distinguishes between tonns that are more than 959%identical)is optional,only being used for the sake g assigned to each independently ae quenced toxin gene,thun deapite the fact that some torins have dsfferent quate Vip Nomenclature; A nomenclature for the Vip and Yp-telaied toxins can be found bere Parasp