白菜RING基因家族的全基因组鉴定与表达分析

发布时间：2018-02-09 11:35

  本文关键词： RING锌指蛋白 生物信息学 非生物胁迫 表达模式 白菜　出处：《福建农林大学》2017年硕士论文　论文类型：学位论文

【摘要】：含有RING结构域的RING指类E3泛素连接酶是参与26S蛋白酶体系统中的最重要的酶类型之一。越来越多的RING指蛋白基因被发现参与到各种重要的生物学过程中。本研究从白菜(Brassica rapa,AA,2n=20)基因组数据中共鉴别出了 715个RING指蛋白基因含731个RING结构域,后者可进一步被分为 8 个类型:RING-H2(371),RING-HCa(215),RING-HCb(47),RING-v(44),RING-C2(38),RING-D(10),RING-S/T(5)和 RING-G(1)。按照是否还含有其它的结构域及其类型,715个RING指蛋白又可被分成51个组。其中700个RING指蛋白基因可以被制图到绘制白菜的10条染色体上:每条染色体上含有47～111个RING指蛋白基因。通过对白菜和拟南芥的RING指蛋白基因之间的同源关系比较显示:在白菜基因组中,仅在50例情况下,被三倍复制的三个拷贝完好的存在于三个亚基因组上(LF,MF1和MF2);而在146例情况下,被三倍复制的三个拷贝中仅有两个拷贝被完好的保留下来;在294例情况下,被三倍复制的三个拷贝中仅有一个拷贝被完好的保留下来。有57个白菜RING指蛋白基因和58个拟南芥RING指蛋白基因发生了衔接重复。RNA-seq转录组数据分析表明:其中有667个RING指蛋白基因在1至多种白菜组织器官中表达,证明它们参与到了白菜的各种生理和发育过程中。通过对它们对应的RNA-seq转录组数据的聚类分析,可以将其分为7个大类,其中1类含有231个都趋于在叶片表达的RING指蛋白基因,因而可以作为研究芸薹属(Brassica)作物叶球性状形成的遗传和分子机制的候选基因。利用半定量RT-PCR技术对其中的29个有代表性的不同类型的RING指蛋白基因进行了表达模式的验证。此外,利用利用实时荧光定量PCR(Quantitative Real-Time PCR,qRT-PCR)技术对其中的 17 个有代表性的RING指蛋白基因在盐和干旱胁迫条件下的表达模式进行了分析,结果发现其中有14和13个基因分别受盐和干旱胁迫诱导表达。本研究结果为进一步研究芸薹属作物的RING指蛋白基因的分类、进化和功能奠定了基础。
[Abstract]:RING finger E3-like ubiquitin ligase containing RING domain is one of the most important enzyme types involved in 26s proteasome system. More and more RING finger protein genes have been found to be involved in various important biological processes. A total of 715 RING finger protein genes containing 731 RING domains were identified from the genome data of Brassica rapa. The latter can be further divided into eight types of RING-H2H2O371CIRING-HCaH215AK, RING-HCbP47, RING-vC2O44-RING-C2C2O4, RING-C2C2O4, RING-C2C2O4, RING-DX, RING-G1T, and RING-G1.There are also 715 RING finger proteins that can be divided into 51 groups according to whether they contain other domains and their types. Of these, 700 RING finger protein genes can be mapped to be plotted in Chinese cabbage (Brassica campestris var. campestris var. campestris). In addition, 700 RING finger protein genes can be mapped and mapped to Chinese cabbage. 10 chromosomes: each chromosome contains 47 ~ 111 RING finger protein genes. The homology of RING finger protein genes between Chinese cabbage and Arabidopsis thaliana shows that: in Chinese cabbage genome, In only 50 cases, three copies of the triple copy were intact in the three subgenomes LFN MF1 and MF2; in 146 cases, only two copies of the triple copy were retained; in 294 cases, Only one of the three copies of the triple copy was intact. 57 RING finger protein genes in Chinese cabbage and 58 Arabidopsis RING finger protein genes were linked to repeat. RNA-seq transcriptome data showed that:. 667 RING finger protein genes were expressed in one or more kinds of Chinese cabbage tissues and organs. It is proved that they are involved in various physiological and developmental processes of Chinese cabbage. By cluster analysis of their corresponding RNA-seq transcriptome data, they can be classified into 7 categories, including 231 RING finger protein genes that tend to express in leaves. Therefore, it can be used as a candidate gene to study the genetic and molecular mechanism of leaf ball character formation in Brassica crops. A semi-quantitative RT-PCR technique was used to express 29 different types of RING finger protein genes in Brassica. In addition, The expression patterns of 17 representative RING finger protein genes under salt and drought stress were analyzed by real-time fluorescence quantitative PCR(Quantitative Real-Time PCR qRT-PCRR. The results showed that 14 and 13 of them were induced by salt and drought stress, respectively, which laid a foundation for further study on the classification, evolution and function of RING finger protein genes in Brassica.
【学位授予单位】：福建农林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S634;Q943.2
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本文编号：1497838