本文關(guān)鍵詞： RING鋅指蛋白 生物信息學(xué) 非生物脅迫 表達(dá)模式 白菜　出處：《福建農(nóng)林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：含有RING結(jié)構(gòu)域的RING指類E3泛素連接酶是參與26S蛋白酶體系統(tǒng)中的最重要的酶類型之一。越來(lái)越多的RING指蛋白基因被發(fā)現(xiàn)參與到各種重要的生物學(xué)過(guò)程中。本研究從白菜(Brassica rapa,AA,2n=20)基因組數(shù)據(jù)中共鑒別出了 715個(gè)RING指蛋白基因含731個(gè)RING結(jié)構(gòu)域,后者可進(jìn)一步被分為 8 個(gè)類型: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)。按照是否還含有其它的結(jié)構(gòu)域及其類型,715個(gè)RING指蛋白又可被分成51個(gè)組。其中700個(gè)RING指蛋白基因可以被制圖到繪制白菜的10條染色體上:每條染色體上含有47～111個(gè)RING指蛋白基因。通過(guò)對(duì)白菜和擬南芥的RING指蛋白基因之間的同源關(guān)系比較顯示:在白菜基因組中,僅在50例情況下,被三倍復(fù)制的三個(gè)拷貝完好的存在于三個(gè)亞基因組上(LF,MF1和MF2);而在146例情況下,被三倍復(fù)制的三個(gè)拷貝中僅有兩個(gè)拷貝被完好的保留下來(lái);在294例情況下,被三倍復(fù)制的三個(gè)拷貝中僅有一個(gè)拷貝被完好的保留下來(lái)。有57個(gè)白菜RING指蛋白基因和58個(gè)擬南芥RING指蛋白基因發(fā)生了銜接重復(fù)。RNA-seq轉(zhuǎn)錄組數(shù)據(jù)分析表明:其中有667個(gè)RING指蛋白基因在1至多種白菜組織器官中表達(dá),證明它們參與到了白菜的各種生理和發(fā)育過(guò)程中。通過(guò)對(duì)它們對(duì)應(yīng)的RNA-seq轉(zhuǎn)錄組數(shù)據(jù)的聚類分析,可以將其分為7個(gè)大類,其中1類含有231個(gè)都趨于在葉片表達(dá)的RING指蛋白基因,因而可以作為研究蕓薹屬(Brassica)作物葉球性狀形成的遺傳和分子機(jī)制的候選基因。利用半定量RT-PCR技術(shù)對(duì)其中的29個(gè)有代表性的不同類型的RING指蛋白基因進(jìn)行了表達(dá)模式的驗(yàn)證。此外,利用利用實(shí)時(shí)熒光定量PCR(Quantitative Real-Time PCR,qRT-PCR)技術(shù)對(duì)其中的 17 個(gè)有代表性的RING指蛋白基因在鹽和干旱脅迫條件下的表達(dá)模式進(jìn)行了分析,結(jié)果發(fā)現(xiàn)其中有14和13個(gè)基因分別受鹽和干旱脅迫誘導(dǎo)表達(dá)。本研究結(jié)果為進(jìn)一步研究蕓薹屬作物的RING指蛋白基因的分類、進(jìn)化和功能奠定了基礎(chǔ)。
[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.
【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S634;Q943.2
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本文編號(hào)：1497838