05月03 【成功案例】百邁客云助力喻達(dá)輝老師 使用 真核生物有參考基因組的轉(zhuǎn)錄組分析平臺 輔助研究，發(fā)表《通過轉(zhuǎn)錄組分析合浦珠母貝對異體血細(xì)胞免疫表達(dá)相關(guān)基因》
摘要：
合浦珠母貝是在中國普遍養(yǎng)殖的海洋珍珠貝，為了培養(yǎng)珍珠，需要將供體珍珠牡蠣的地幔片用細(xì)胞核移植到受體中，而這個移植的幔片可能會被受體貝珍珠貝的免疫系統(tǒng)所攔截，從而降低移植成功率，然而，現(xiàn)在關(guān)于珍珠貝對同種異體移植的免疫防御報道很少。在本研究中，對同種異體移植后0小時和48小時的珍珠貝的血細(xì)胞免疫反應(yīng)進(jìn)行了轉(zhuǎn)錄組分析。測序得到92.5M的clean reads，與合浦珠母貝的參考基因組進(jìn)行了比對，為了鑒定所有與免疫相關(guān)的差異表達(dá)基因，本研究進(jìn)行了GO注釋和KEGG通路分析。與0小時相比，在移植后48個小時的樣本中總共有798個差異表達(dá)基因（410個上調(diào)基因，388個下調(diào)基因），其中血細(xì)胞中白細(xì)胞介素受體和toll-like受體的表達(dá)水平顯著升高，表明移植誘導(dǎo)了珍珠貝的免疫應(yīng)答反應(yīng)。最后，本研究對其中的18個隨機(jī)選取的免疫相關(guān)的差異表達(dá)基因進(jìn)行了qRT-PCR驗證。本文的研究結(jié)果為進(jìn)一步分析同種異體移植的免疫排斥提供了依據(jù)。
本文主要用了百邁客云平臺主流程。
英文摘要：
The pearl oyster Pinctada fucata is commonly cultured for marine pearls in China. To culture pearls, a mantle piece from a donor pearl oyster is grafted with a nucleus into a receptor. This transplanted mantle piece may be rejected by the immune system of the recipient oyster, thus reducing the success of transplantation. However, there have been limited studies about the oyster’s immune defense against allograft. In this study, hemocyte transcriptome analysis was performed to detect the immune responses to allograft in P. fucata at 0 h and 48 h after a transplant. The sequencing reaction produced 92.5 million reads that were mapped against the reference genome sequences of P. fucata. The Gene Ontology (GO) annotation and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to identify all immune- related differentially expressed genes (DEGs). Compared with patterns at 0 h, a total of 798 DEGs were identified, including 410 up-regulated and 388 down-regulated genes at 48 h. The expression levels of interleukin receptor and toll-like receptor in hemocytes were increased significantly 48 h post- transplant, indicating that the oyster immune response was induced. Finally, altered levels of 18 randomly selected immune-related DEGs were confirmed by quantitative real-time PCR (qRT-PCR). Our results provide the basis for further analysis of the immune rejection of allotransplantation.