• 作者： 歐樂君; 黃裕斌; 鄭後琴; 黃德富; 朱廣邦
• 作者服務機構： 臺北榮民總醫院醫學研究部; 國立陽明醫學院遺傳研究部; 國立臺灣大學藥理研究所
• 中文摘要： 利用 －噬菌體的cDNA基因庫，已發展出可迅速篩選和確認cDNA的選殖體的技術。此法是由部份蛋白質氨基酸序列
  合成一混合鹼基寡核?酸為探針，進行雜交篩選，所獲得的cDNA選殖體利用載體上下游之端子，以聚合?連鎖反應法
  (PCR)擴增此片段，所得的片段再殖入質體中，同時，利用混合寡核?酸為定序端子進行核?酸序列之鑑定。利用此法成
  功地篩選出部份基因密碼已知的血小板凝聚抑制因子之選殖體。
  e
• 英文摘要： A rapid procedure has been developed for the isolation and verification of cDNA clones isolatedfrom a cDNA library based on lambda vectors. Using information of the partial amino acid sequence of aprotein, synthetic mixed-base oligonucleotides are first employed as a screening probe using the plaquehybridization procedure. The cDNA inserts of the clones obtained are then directly amplified by poly-merase chain reaction (PCR) using primers flanking the cloning site of the vector. Besides being used forcloning into a plasmid vector, the amplified DNA's are also subjected to nucleotide sequence analysisusing the same mixed-base oligonucleotides as sequencing primers. This approach allows sequencingthrough the region of the known amino acid sequence for direct verification of the authenticity of theclones obtained. This procedure has successfully been used for cloning and partial characterization of thegene coding for a platelet aggregation inhibitor.
• 中文關鍵字： cDNA library; oligonucleotied primer; nucleotide sequencing
• 英文關鍵字： --