本文選題：鱸魚 + CDγ/δ��； 參考：《農(nóng)業(yè)生物技術(shù)學(xué)報(bào)》2016年03期

【摘要】：T淋巴細(xì)胞分化抗原3(cluster of differentiation 3,CD3)分子表達(dá)于T細(xì)胞表面,并通過非共價(jià)鍵與T細(xì)胞受體(T cell receptor,TCR)連接,參與T細(xì)胞活化。本研究采用反轉(zhuǎn)錄PCR(RT-PCR)和c DNA末端快速擴(kuò)增(rapid amplification of c DNA ends,RACE)技術(shù)從鱸魚(Lateolabrax japonicus)肝臟中克隆了CD3γ/δ、CD3ε和CD3ζ基因全長(zhǎng)。結(jié)果表明,CD3γ/δ全長(zhǎng)c DNA為1 231 bp,包括99 bp的5'非翻譯區(qū)(untranslated regions,UTR),583 bp的3'-UTR和549 bp的開放閱讀框(open reading frame,ORF),共編碼182個(gè)氨基酸;CD3ε全長(zhǎng)c DNA為2 145 bp,包括186 bp的5'-UTR,1 434 bp的3'-UTR和525 bp的ORF,共編碼174個(gè)氨基酸;CD3ζ全長(zhǎng)1 281 bp,包括56 bp的5'-UTR,772 bp的3'-UTR和453 bp的ORF,共編碼150個(gè)氨基酸。氨基酸序列分析發(fā)現(xiàn),CD3γ/δ和CD3ε分子結(jié)構(gòu)相似,均含有1個(gè)免疫球蛋白樣結(jié)構(gòu)的胞外區(qū)、1個(gè)跨膜區(qū)和含1個(gè)免疫受體酪氨酸激活基序(Immunoreceptor tyrosine-based activation motif,ITAM)的胞漿區(qū),而CD3ζ含有1個(gè)僅由5個(gè)氨基酸組成的胞外區(qū)、1個(gè)跨膜區(qū)和含3個(gè)ITAM序列的胞漿區(qū)。分析DNA和c DNA序列發(fā)現(xiàn),CD3γ/δ的ORF區(qū)由6個(gè)外顯子和5個(gè)內(nèi)含子組成,CD3ε的ORF區(qū)由5個(gè)外顯子和4個(gè)內(nèi)含子組成。q RT-PCR結(jié)果表明,CD3γ/δ、CD3ε和CD3ζ在鰓、脾、頭腎、腸中表達(dá)較高,在脂肪、肝、眼、腦等組織中表達(dá)較少。當(dāng)腹腔注射哈維氏弧菌(Vibrio harveyi)3 h后,脾組織的3種CD3分子均顯著上調(diào)(P0.05),在腸和頭腎也有顯著上調(diào)(P0.05),表明細(xì)菌感染會(huì)增加CD3的表達(dá)。研究結(jié)果為進(jìn)一步研究魚類CD3在病原菌感染免疫中的分子機(jī)制提供基礎(chǔ)資料。
[Abstract]:T-lymphocyte differentiation antigen 3 (cluster of differentiation 3 + CD3) is expressed on the surface of T cells and is associated with T cell receptor (T cell receptor) via non-covalent bonds. In this study, the full length of CD3 緯 / 未 T CD3 蔚 and CD3 味 genes were cloned from the liver of Lateolabrax japonicus by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of (rapid amplification of c DNA endsrace. The results show that the full-length cDNA of CD3 緯 / 未 is 1 231 BP, including a untranslated region of 99 BP (583 BP) and an open reading frame (open reading frame ORF) of 583 BP. The full-length c DNA encoding 182 amino acid sequence CD3 蔚 is 2 145 BP, including 186 BP of 5 '-UTR1 434 BP of 3FT-UTR and 525 BP of BP. ORF, encoding 174 amino acids CD3 味 with a total length of 1 281 BP, including 56 BP 5- UTR772 BP 3- UTR772 BP 3-UTR and 453-BP ORF, encoding a total of 150 amino acids. Amino acid sequence analysis showed that the molecular structures of CD3 緯 / 未 and CD3 蔚 were similar, with one extracellular region with immunoglobulin-like structure, one transmembrane region and one cytoplasmic domain containing immune receptor tyrosine-based activation motifitm. CD3 味 contained an extracellular domain consisting of only 5 amino acids, a transmembrane region and a cytoplasmic domain containing three ITAM sequences. The ORF region of CD3 緯 / 未 was composed of 6 exons and 5 introns. Q RT-PCR showed that CD3 緯 / 未 CD3 蔚 and CD3 味 were expressed in Gill, spleen, head kidney, intestine, fat, liver and liver. Eye, brain and other tissues are less expressed. After intraperitoneal injection of Vibrio harveyi for 3 h, three kinds of CD3 molecules in spleen tissue were significantly up-regulated (P0.05), as well as in intestine and cephalic kidney (P0.05), indicating that bacterial infection could increase the expression of CD3. The results provide basic data for further study of the molecular mechanism of fish CD3 in pathogen infection immunity.
【作者單位】： 寧波大學(xué)海洋學(xué)院;
【基金】：浙江省科技廳創(chuàng)新團(tuán)隊(duì)項(xiàng)目(No.2012R10029-08) 寧波大學(xué)“水產(chǎn)”浙江省重中之重開放基金資助(xkzsc1513)
【分類號(hào)】：S917.4
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本文編號(hào)：2061237