黑鯛第一型類胰島素生長因子基因片段之合成

CNU IR > Chna Nan Annual Bulletin > Vol.20 (1994) > Item 310902800/23173

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Title:	黑鯛第一型類胰島素生長因子基因片段之合成 Amplification of Black Seabream Insulin-Like Growth Factor I Dna by Polymerase Chain Reaction
Authors:	吳鴻程范晉嘉陳泓志張繼堯吳金洌
Contributors:	食品衛生科
Keywords:	黑鯛核苷聚合酶連鎖反應類胰島素生長因子 DNA核苷引子 black seabream polymerase chain reaction isulin-like growth factor I DNA primer
Date:	1994
Issue Date:	2010-11-10 18:22:10 (UTC+8)
Abstract:	依據黑鯛第一型類胰島素生長因子（Insulin-like growth factor I, IGF-1）DNA序列，合成長度為23核苷酸的S核苷引子，19個核苷酸的B核苷引子，21個核苷酸的D核苷引子和22個核苷酸的E核苷引子。配對後的核苷引子利用Vent核苷聚合酶進行連鎖反應；其反應條件為：解鏈溫度94℃（反應1分鐘），黏合溫度65℃（反應2分鐘），合成溫度72℃（反應2分鐘），共重複35次循環。經核苷酸聚合酶連鎖反應後，可生成B區域到D區域201個核苷酸DNA片段；B區域到E區域423個核苷酸的DNA片段，S區域到E區域552個核苷酸的DNA片段。S區域到D區域的DNA片段，則在解鏈溫度94℃(反應30秒)，黏合溫度54℃ (反應30秒)，合成溫度74℃(反應30秒)，共進行35次循環的條件下可生成330個核背酸之DNA片段。 Four oligonuclotide primers, primer S (23 bases), primeer B (19 bases), primer D (21 bases)and primer E (22 bases), were synthesized according to the black seabream (Acanthopagrus schegeli) insulin-like growth factor I (IGF-1) DNA sequence to amplify the IGF-1 DNA fragments.The results found that primeer S, primer E produced 552 base pairs IGF-1 DNA fragment, primer B, primer D producted 201 base pairs IGF-1 DNA fragment, and primer B, primer D produced 423 base pairs IGF-1 fragment at denatured 1 minute at 94℃, annealized 2 minutes at 65℃, and extended 2 minutes at 72℃ by polymerase chain reaction. Primer S and primer D producted 330 base pairs JGF-1 DNA fragment at denatured 30 seconds at 94℃, annealized 30 seconds at 54℃ and extended 30 seconds at 74℃ by polymerase chain reaction.
Relation:	嘉南學報 20期：p.138-147
Appears in Collections:	[Chna Nan Annual Bulletin] Vol.20 (1994) [Dept. of Food Science & Technology] Periodical Articles

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