本文選題：睪丸 + 綿羊�。� 參考：《農(nóng)業(yè)生物技術學報》2017年05期

【摘要】：本研究旨在克隆綿羊(Ovis aires)視黃醇結合蛋白4(retinol binding protein 4,RBP4)基因的CDS區(qū)序列,檢測其在綿羊不同月齡睪丸組織中的表達水平,從而為研究RBP4基因在綿羊睪丸發(fā)育中的作用奠定基礎。以綿羊睪丸為材料,采用RT-PCR技術克隆RBP4基因并用相關生物信息學軟件預測RBP4蛋白的結構,采用qRT-PCR和免疫組織化學技術分別檢測RBP4 m RNA和蛋白質的表達水平。結果顯示,克隆得到綿羊RBP4基因CDS區(qū)長度為606 bp,編碼201個氨基酸。綿羊RBP4核苷酸序列與山羊(Capra hircus)、牛(Bos taurus)、豬(Sus scrofa)、人(Homo sapiens)和小鼠(Mus musculus)的相似性分別為97%、97%、95%、91%和85%。綿羊RBP4基因與山羊具有最近的親緣關系,其次為牛、豬、人和小鼠。RBP4 m RNA在不同月齡綿羊睪丸中均有表達,表達量隨月齡增加呈現(xiàn)出先降低后上升的趨勢,相鄰兩組之間差異極顯著(P0.01)。RBP4蛋白主要表達于初級精母細胞、精子細胞、精子和生精小管,陽性反應積分光密度值隨月齡增加呈現(xiàn)出先升高后降低的趨勢,與綿羊睪丸RBP4 m RNA表達量相反。本研究結果表明,RBP4基因可能參與綿羊睪丸發(fā)育的調控,為進一步探討RBP4在綿羊生殖過程中發(fā)揮的作用提供參考資料。
[Abstract]:The aim of this study was to clone the CDS region of the retinol binding protein 4 (RBP4) gene of sheep (vis aires) and detect its expression level in the testis of sheep at different months of age, so as to lay a foundation for the study of the role of RBP4 gene in the development of sheep testis. RBP4 gene was cloned from sheep testis by RT-PCR and the structure of RBP4 protein was predicted by bioinformatics software. The expression levels of RBP4 mRNA and protein were detected by qRT-PCR and immunohistochemistry, respectively. The results showed that the length of CDS region of sheep RBP4 gene was 606 BP, encoding 201 amino acids. The similarity between sheep RBP4 nucleotide sequence and goat (Bos taurus), pig (Sus scrofa), human (Homo sapiens) and mouse (Mus musculus) was 97,9795% and 85.1%, respectively. Sheep RBP4 gene was closely related to goat, followed by bovine, pig, human and mouse. RBP4 mRNA was expressed in testis of sheep at different months of age. The expression of RBP4 mRNA decreased first and then increased with the increase of age. The difference between the two groups was significant (P0.01). RBP4 protein was mainly expressed in primary spermatocytes, spermatozoa and seminiferous tubules. The integral optical density of positive reaction increased first and then decreased with the increase of age. The expression of RBP4 mRNA was opposite to that of sheep testis. The results suggest that RBP4 gene may be involved in the regulation of testis development in sheep, and provide references for further study of the role of RBP4 in sheep reproductive process.
【作者單位】： 甘肅農(nóng)業(yè)大學動物科學技術學院;甘肅肉羊繁育生物技術工程實驗室;甘肅農(nóng)業(yè)大學動物醫(yī)學院;
【基金】：甘肅省科技重大專項計劃(No.1602NKDH020) 甘肅農(nóng)業(yè)大學校級課題(No.GSAU-ZL-2015-030) 甘肅省農(nóng)業(yè)生物技術研究與應用開發(fā)項目(No.GNSW-2012-19) 國家肉羊產(chǎn)業(yè)技術體系(No.CARS-39)
【分類號】：Q78;S826

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