阿拉伯芥突變種(hit1)之位址定位; Genetic Mapping of hit1 Locus in Arabidopsis thaliana

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题名:	阿拉伯芥突變種(hit1)之位址定位;Genetic Mapping of hit1 Locus in Arabidopsis thaliana
作者:	蒲欣儀;Shin-Yi Pu
贡献者:	生命科學研究所
关键词:	阿拉伯芥;對熱不耐受性突變株;hit1;heat intolerance;Arabidopsis
日期:	2004-07-08
上传时间:	2009-09-22 10:17:44 (UTC+8)
出版者:	國立中央大學圖書館
摘要:	中文摘要為了了解植物耐受高溫的遺傳控制因子及其生理機制，我們用化學突變劑( EMS )誘導阿拉伯芥點突變產生，並進而篩選出一株對熱缺乏耐受性之突變種，稱之為hit1( heat intolerance ) 。這是一種典型從具遺傳性之生理性狀找出相對應之基因及其分子作用的研究方式，亦即所謂的向前式遺傳學分析法( forward genetics )，為功能性遺傳學研究法中的一種。因 hit1 之性狀為點突變所造成的，故其功能性基因須以遺傳圖譜為基礎來進行定位選殖( map-based cloning, MBC ) 。其做法是先將Columbia 生態型之hit1 突變種與Landsberg 生態型之野生種雜交，之後的F1 自交得F2 世代，F2 世代中帶有hit1 性狀之個體為hit1 定位時所需之分離族群 ( segregation population )。以hit1 之突變性狀，37℃處理四天後會死亡的原則篩選，再經簡單序列長度多型性( simple sequence length polymorphism , SSLP )及限制酵素切割擴增片段多型性( cleaved amplified polymorphic sequence , CAPS )兩種對生態型具專一性之分子標記進行定位。本研究共設計了26 組SSLP 分子標記，7 組CAPS 分子標記。在檢測過3987 株F2 世代的hit1 突變種之後，推得hit1 基因坐落在第一條染色體， AGI map 18767.3 kb ~ 18810.2 kb 之42.9kb 範圍內，達到MBC 所需之定位目標。 Abstract In order to understand the mechanism of plant crop heat stress, we screened a mutant that is difficult to tolerant heat stress called “hit1” (heat intolerance) inducible by chemical mutagenesis, EMS, causing a point mutation. Forward genetic is one of functional genetics approaches, and it is underlying a mutant with a desirable genetic phenotype to an identified mutation in a gene and to analysis specific gene’s molecular biology function. We use forward genetic to approach “hit1” that was a point mutation, so we want a cloning of sequencing mutant-define gene by map-based coloning (MBC). A hit1 mutant with Columbia ecotype (P1) and a wild type with Landsberg erect ecotype (P2) were parent to get recombinant inbred lines (F2). It has hit1 mutant specific phenotype lines of F2 generation that is a segregation population to provide hit1 genetic mapping. The definition of hit1 mutant phenotype was death at 37℃ in 4 days to screen mutants from F2 generation as a segregation population then make hit1 gene genetic mapping, and we designed 26 pairs primers of simple sequence length polymorphism (SSLP) markers and 7 pairs primers of cleavage amplify polymorphisms (CAPS) markers for genetic analysis. We suggest that hit1 gene locus in AGI 18767.3kb ~ 18810.2kb, total 42.9kb regions, and approach the fine-scale mapping of MBC.
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