冷誘導條件下番茄cDNA文庫的構建及SlNAC29的克隆與表達分析

摘要：低溫是限制冷敏感植物產(chǎn)量和地理分布的重要因素。冷害嚴重影響番茄的產(chǎn)量和品質(zhì)。因此，研究并提高其抗寒性具有重要的理論和現(xiàn)實意義。本研究將番茄幼苗置于4℃下處理，取其葉片為試材，構建了差減cDNA文庫，為番茄重要抗寒基因的挖掘奠定了基礎。分析冷脅迫下基因表達情況，共獲得91個差異表達基因。根據(jù)文庫信息，利用RT-PCR技術從冷處理的番茄葉片中克隆到一個NAC基因。該基因全長為1243 bp，包括828 bp的開放閱讀框，編碼276 個氨基酸。通過qRT-PCR分析，該基因受低溫及高溫誘導表達。
關鍵字：cDNA文庫，番茄，SlNAC29，低溫


Construction of a tomato cDNA library induced by cold and cloning and expression analysis of SlNAC29

Abstract：Low temperature is the major factor limiting the productivity and geographical distribution of chilling-sensitive plant species. Therefore, it was theoretically and practically significant to study and enhance the cold-tolerance in plants. A subtractive cDNA library of Lycopersicon esculentum Mill was constructed using the 4℃ treated leaves, lying the foundation for selecting and cloning tomato cold- tolerance gene. The expression pattern of cold related gene was analyzed and 91 genes which were differently expressed were obtained. Furthermore, one NAC gene was cloned from the library. The length of this gene was 1243bp, containing a ORF of 828 bp. This gene encoded a protein of 276 amino acids. By analyzing of qRT-PCR, this gene was induced expression by low temperature and high temperature.
Key words：cDNA library, tomato, SlNAC29, low temperature

目錄
摘要····································································································1
關鍵字·································································································1
1前言··································································································2
1.1 cDNA文庫····················································································2
1.1.1 cDNA文庫的特點··········································································2
1.1.2 cDNA文庫構建方法···································································2
1.2 NAC轉錄因子家族··········································································3
1.2.1 NAC轉錄因子的起源·································································3
1.2.2 NAC轉錄因子的結構與分類························································3
1.2.3 NAC轉錄因子的生物學功能························································5
1.2.3.1影響植物的生長發(fā)育·····························································5
1.2.3.2參與多種植物逆境脅迫防御反應··············································5
1.2.4 NAC轉錄因子的表達調(diào)控···························································6
1.3 主要技術路線···············································································6
2 材料與方法························································································6
2.1 試驗材料與處理············································································6
2.1.1 植物材料················································································6
2.1.2 材料處理················································································6
2.1.3菌株與載體··············································································7
2.1.4酶及生化試劑································································..