主持十二五國家863子課題“海洋動(dòng)物配子和胚胎冷凍保存及種質(zhì)庫信息化管理技術(shù)研究”,國家自然科學(xué)基金面上項(xiàng)目“基于轉(zhuǎn)錄組和microRNA 信息的牙鲆白化發(fā)生機(jī)制研究”,國家自然科學(xué)基金青年基金項(xiàng)目“大菱鲆Grim19基因在細(xì)胞凋亡及病原感染防御中的作用研究”,中國水產(chǎn)科學(xué)研究院基本科研業(yè)務(wù)費(fèi)項(xiàng)目“半滑舌鰨雌性特異基因的表達(dá)及功能分析”等。作為科研骨干參加公益性行業(yè)科研專項(xiàng)、國家自然科學(xué)基金重點(diǎn)項(xiàng)目、國家十一五、十二五863項(xiàng)目等10余項(xiàng)。 1)建立并分析了多種鲆鰈魚類細(xì)胞系,如大菱鲆腎細(xì)胞系、漠斑牙鲆腎細(xì)胞系、半滑舌鰨精巢細(xì)胞系、半滑舌鰨頭腎細(xì)胞系、半滑舌鰨卵巢細(xì)胞系等,揭示這些細(xì)胞系在研究魚類病原學(xué)和基因功能方面具有重要的應(yīng)用價(jià)值,其中大菱鲆腎臟細(xì)胞系已作為一個(gè)極為重要的實(shí)驗(yàn)平臺(tái),被國家海洋局第一海洋研究所、上海海洋大學(xué)等多家科研單位成功應(yīng)用于RNA干擾、亞細(xì)胞定位、蛋白生物活性、基因互作等多項(xiàng)研究中。 2)針對(duì)大菱鲆免疫抗病相關(guān)基因GRIM-19,STAT2,STAT3、HSC70、Hepcidin等,開展了免疫應(yīng)答、亞細(xì)胞定位、細(xì)胞凋亡等研究,揭示了它們?cè)诘挚滚牷【土馨湍夷[病毒感染方面的重要作用,初步確認(rèn)了GRIM-19和STAT3在大菱鲆細(xì)胞系中的共定位關(guān)系,鑒定了STAT2的核輸出信號(hào),這些基因的鑒定與功能分析為鲆鰈魚類抗病分子育種策略的制定以及抗病功能基因產(chǎn)品的研制提供了基礎(chǔ)資料。 3)闡明了半滑舌鰨雌性特異候選基因CSW3,CSW4等的時(shí)空表達(dá)模式,分析了不同結(jié)構(gòu)域的亞細(xì)胞定位信息,鑒定了重組表達(dá)蛋白的生物學(xué)活性,構(gòu)建了TALEN敲除載體并在模式魚青鳉中進(jìn)行了敲除效率的驗(yàn)證。 4)開展了牙鲆白化相關(guān)的轉(zhuǎn)錄組及miRNA高通量測(cè)序分析,初步篩選獲得差異表達(dá)基因235個(gè),差異表達(dá)miRNA46個(gè),針對(duì)這些相關(guān)基因/miRNA進(jìn)行了GO及KEGG通路富集分析,為牙鲆白化分子機(jī)制的闡示提供了豐富的分子基礎(chǔ)。 目前已在Dev Comp Immunol、Fish Shellfish Immunol、J Fish Dis等刊物上發(fā)表論文20多篇,其中第一作者SCI文章5篇,作為第一發(fā)明人獲授權(quán)專利2項(xiàng),參編專著1部。
Dr Wang conducted one National Science Foundation for Young Scholars of China, one National Science Foundation of China, one 863 sub-project, and one Special Scientific Research Funds for Central Non-profit Institutes, Chinese Academy of Fishery Sciences. Also, Dr Wang participated in several projects including 973, 863, the State Key Program of National Natural Science of China, Special Fund for Agro-scientific Research in the Public Interest. 1) Dr Wang established and identified several flatfish cell lines, such as turbot kidney cell line, Southern flounder cell line, half smooth tongue sole testis cell line, and half smooth tongue sole head kidney cell line. Their application values in the fish pathogenesis and gene function were also analyzed. Especially, turbot kidney cell line has been used for the study of RNA interference, sub-cellular location, protein activity, gene interaction and so on. 2) The turbot immune related genes GRIM-19, STAT2, STAT3, HSC70 and Hepcidin were cloned and analyzed, and their possible roles against pathogen affection were revealed. The identification and functional analysis of these genes provide comprehensive basis for molecular disease-resistant breeding in aquaculture. 3) The female specific genes CSW3 and CSW4 of Half smooth tongue sole were studied by expression pattern analysis, recombinant proteins identification and TALEN analysis. 4) The transcriptome and miRNA expression pattern between albinism and normal Japanese flounder are analyzed by high-throughput sequencing. There are 235 differentially expressed genes and 46 differentially expressed miRNAs identified. Further GO and KEGG pathway enrichment were also conducted. These studies provide abundant molecular basis for exploring the mechanism of flatfish albinism. Dr Wang has published over 20 papers in Journals such as Dev Comp Immunol, Fish Shellfish Immunol, J Fish Dis and so on, of which 5 SCI papers were published as the first author. As well, as the first executers, Dr Wang also obtained two authorized patents. |