i基因文庫(kù)與篩選ppt課件(參考版)
2025-05-01 22:05本頁(yè)面
【正文】 。 南京林業(yè)大學(xué) 扣除雜交（ Subtractive Hybridization） 南京林業(yè)大學(xué) Section I: Gene libraries and screening Hybrid arrest (雜交扣留 ) Hybrid arrested translation: cDNA?hybridized mRNA ? proteins ? Hybridization: Individual cDNA clones or pools (群 ) of clones can be hybridized to mRNA preparations. The hybridized mRNA can not be translated to proteins, and the detection of proteins can be used to find which cDNA can arrest which protein. ? Subdivision (細(xì) 分 ): of the pools of cDNA into smaller numbers and repeating the experiment should allow the identification of a single cDNA that arrests (扣留 ) the translation of one protein. mRNA cDNA 應(yīng)用細(xì)胞外翻譯系統(tǒng) 南京林業(yè)大學(xué) Section I: Gene libraries and screening Hybrid release (雜交釋放 ) Hybrid release translation: cDNA?hybridized mRNA?proteins ? Purification: After hybridization, the hybrids can be purified, . (例如 ) the [cDNA/mRNA] are attached to magic beads or are precipitable (凝集 ) with an antibody on membrance. ? Release: Then the mRNAs are released from hybrids (by heat or denaturant) and translated (翻譯 ). This hybrid release translation identifies which cDNA clone encode which protein. mRNA mRNA cDNA 南京林業(yè)大學(xué) 染色體步移 從第一個(gè)重組克隆插入片段的一端分離出一個(gè)片段作為探針篩選第二個(gè)重組克隆，該克隆插入片段含有與探針重疊順序和染色體的其他順序。為了進(jìn)一步提高差別雜交的篩選效率，一種切實(shí)可行的辦法是 構(gòu)建富含目的基因序列的 cDNA文庫(kù) 。 南京林業(yè)大學(xué) 南京林業(yè)大學(xué) 南京林業(yè)大學(xué) 南京林業(yè)大學(xué)
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