NF-Y转录因子标签表达载体的构建开题报告

1. 研究目的与意义、国内外研究现状（文献综述）

1. 研究背景

1. NF-Y转录因子在植物发育中的重要作用

在植物中，每个 NF-Y 亚基都是由1个多基因家族编码的。 NF-Y 转录因子的基因家族基因的亚基类型有 NF-YA 、 NF-YB 和 NF-YC 。例如在模式植物拟南芥中， NF-Y 家族成员众多，包括10个 NF-YA 、13个 NF-YB 和13个 NF-YC 。NF-Y亚基，在不同的植物中，生物学功能大致相同，主要是控制植物在生长发育过程中对在胚胎和种子发育有促进作用，在调控植物开花中有促进作用，在非生物胁迫反应中对植物的抗性有增强 (宋秋明等，2015) 。

(1.)在不同物种中 NF-YA 的生物学功能如下：

(2).在不同物种中 NF-YB 的生物学功能如下:

(3).在不同物种中NF-YB的生物学功能如下:

目前仅限于在模式植物拟南芥和一些具有基因组信息的植物上做了一些简单的生物信息学比对和初步的功能研究。上述图表 (丁慧霞 等，2017) 表明 NF-Y 的功能十分广泛，尤其是在逆境响应和生长发育方面，为人们认识植物中 NF-Y 家族提供了大量的基础数据。

2. 用ChIP-seq技术研究转录因子全基因组结合位点

ChIP-seq原理:首先通过染色质免疫共沉淀技术特异性地富集目的蛋白结合的DNA片段，并对其进行纯化与文库构建；然后对富集得到的DNA片段进行高通量测序。通过将获得的数百万条序列标签精确定位到基因组上，从而获得全基因组范围内与组蛋白、转录因子等互作的DNA区段信息。

ChIP-seq先通过ChIP技术特异性地富集能与目的蛋白结合的DNA然后对这些DNA高通量测序，测序后首先需要将序列比对到已知基因组上并确立真正的结合位点。对于转录因子, 要寻找与“峰”游调控基因, 或者构建转录因子结合位点的保守结合序列(Motif); 如果转录因子的motif是已知的, 则可以计算“峰”序列中包含 motif 的序列百分比, 间接估计实验结果的可靠性。ChIP-seq是研究转录因子全基因组结合位点的强力工具，已经对例如AP2等多个重要的基因展开了相关研究。

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2. 研究的基本内容和问题

2.研究目的：

要研究NF-Y的调控，做ChIP-seq的前提是需要好的抗体，本实验是将NF-Y转录因子与标签融合，可利用标签的抗体进行NF-Y的ChIP实验。

3. 研究的方法与方案

2.实验的可行性：

2.1本次试验知识基础牢固、方案设计可行

本人已完成分子生物学、专业文献阅读、专业文献写作，已自主学习了分子生物学以及阅读了大量有关nf-y转录因子、chip-seq的分析流程的文献积累了一定的对于本课题的知识，而且对本课题十分感兴趣，积极性比较高。

4. 研究创新点

5. 研究计划与进展

4．研究计划及预期进展

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