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【正文】 btractive Hybridization） 南京林業(yè)大學 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 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)用細胞外翻譯系統(tǒng) 南京林業(yè)大學 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 染色體步移 從第一個重組克隆插入片段的一端分離出一個片段作為探針篩選第二個重組克隆，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。 從第二個重組克隆的插入片段再分離出末端小片段篩選第三個重組克隆，如此重復，得到一個相鄰的片段，等于在染色體上移了一步 . 是一種重要的分子生物學研究技術(shù)，使用這種技術(shù)可以有效獲取與已知序列相鄰的未知序列。
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