• 作者： 陳建華; 葉修足
• 作者服務機構： 中興大學分子生物研究所
• 中文摘要： 大腸桿菌K-12品系染色體上已知有六套IS1(IS1A-IS1F)。依據其核?酸序列，可分成四種：IS1A(IS1E)，
  IS1B(IS1C），IS1D，IS1F。但Zuber與Schumann於1993年進一步於W3110品系的49.6分鐘位置又發現有第七套IS1
  的存在，並且發表此套IS1尾端21-bp及其外圍約120-bp的序列。我們根據IS1A(IS1E)，IS1B(IS1C)，IS1D的前端16
  鹼基序列及第七套IS1尾端外圍18鹼基的序列合成二個引子，將W3110染色體上第七套is]以聚合?連鎖反應的方法
  複製出來，並加以定序。結果顯示第七套IS1序列與IS1A(IS1E)序列完全相同。我們進一步利用一質體系統篩選染色體
  IS轉位到質體的質體自發性突變株，總共得到142株質體IS轉位株。這其中有38株為IS5之轉位株，10株為IS30之
  轉位株，而其餘94株為IS1之轉位株。以限制脢圖譜分析此94株IS1轉位株，我們發現其皆為具有IS1A(IS1E)序列的
  IS1轉位株。
  ：
• 英文摘要： In the Escherichia coli K-12 chromosome, six copies of IS1 (IS1A - IS1F) have been identified andcharacterized. According to their nucleotide (nt) sequences, the six IS1 copies can be classified into fourtypes, IS1A(IS1E), IS1B(IS1C), IS1D and IS1F type. Zuber and Schumann (1993) identified the seventhIS1 copy at 49.6 minutes on the E. coli W3110 genetic map. Unfortunately, only the end 21-bp seauence aswell as the neighboring 120-bp E. coli sequence were reported. we therefore designea two ohgonucieotiaeprimers to specifically amplify the seventh IS1 copy by PCR. One primer is homologous to the first sixteenbases of the IRL sequence of IS1A(IS1E)，IS1B(IS1C) and IS1D. The other primer is complementary to theeighteen bases of E. coli sequence adjacent to IRR of the seventh IS1 copy. An 800-bp PCR fragment wasobtained and its nt sequence determined, revealing an identical nucleotide sequence to that of IS1 A(IS1E).A plasmid system was then used to isolate insertion mutations caused by insertions of the chromosomal inser-tion sequences. Of the 142 plasmid insertion mutants isolated, thirty-eight were insertions of chromosomalIS5, ten were IS30, and ninety-four were IS1. Detailed restriction mapping indicates that all ninety-fourplasmid IS1 insertions were insertions of IS1 of the IS1A(IS1E) type.
• 中文關鍵字： IS1; PCR; transposition.
• 英文關鍵字： --