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利用实时定量逆转录聚合酶链反应分析爪哇根结线虫分支酸变位酶转录本表达谱

Meloidogyne javanica Chorismate Mutase Transcript Expression Profile Using Real-Time Quantitative RT-PCR.

作者信息

Painter Janet E, Lambert Kris N

出版信息

J Nematol. 2003 Mar;35(1):82-7.

Abstract

A developmental expression profile of the Meloidodgyne javanica esophageal gland gene chorismate mutase-1 (Mj-cm-1) could suggest when in the lifecycle of the nematode the Mj-cm-1 product is functional. This study used real-time quantitative RT-PCR to examine the variation in Mj-cm-1 transcript levels over six timepoints in the nematode lifecycle: egg, infective second-stage juveniles (Inf-J2), 2-day post-inoculation (pi), 7-day pi, 14-day pi, and adult. The Mj-cm-1 mRNA levels peaked at 2-day pi, about 100-fold above levels expressed at the egg and Inf-J2 stages. Some expression of Mj-cm-1 remained during the 7-day pi, 14-day pi, and adult stages. High transcript levels of the beta-actin control gene M. javanica Beta-actin-1 (Mj-ba-1) demonstrated the presence of cDNA at all timepoints. The peak in Mj-cm-1 transcript expression at 2-day pi as well as the previously shown esophageal gland localization of Mj-cm-1 mRNA suggest that the product of this gene may be involved early in the establishment of parasitism.

摘要

爪哇根结线虫食管腺基因分支酸变位酶-1(Mj-cm-1)的发育表达谱,可能提示该线虫生命周期中Mj-cm-1产物何时具有功能。本研究采用实时定量RT-PCR检测线虫生命周期六个时间点(卵、感染性二期幼虫(Inf-J2)、接种后2天(pi)、接种后7天、接种后14天和成虫)Mj-cm-1转录水平的变化。Mj-cm-1 mRNA水平在接种后2天达到峰值,比卵期和Inf-J2期表达水平高约100倍。在接种后7天、接种后14天和成虫期,Mj-cm-1仍有一些表达。β-肌动蛋白对照基因爪哇根结线虫β-肌动蛋白-1(Mj-ba-1)的高转录水平表明在所有时间点均存在cDNA。接种后2天Mj-cm-1转录本表达峰值以及之前显示的Mj-cm-1 mRNA食管腺定位表明,该基因产物可能在寄生关系建立早期发挥作用。

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