本文選題：梅花鹿 + 胸腺素beta�。� 參考：《生物技術(shù)通報(bào)》2017年06期

【摘要】：轉(zhuǎn)錄組測(cè)序發(fā)現(xiàn)Tβ10在鹿茸中高表達(dá),為了研究Tβ10與鹿茸生長(zhǎng)發(fā)育間的關(guān)系,構(gòu)建Tβ10真核表達(dá)載體。使用Trizol法提取梅花鹿鹿茸總RNA,PCR技術(shù)特異性擴(kuò)增梅花鹿Tβ10基因,利用酶切位點(diǎn)將目的片段插入真核表達(dá)載體VR1012構(gòu)建表達(dá)質(zhì)粒,通過Fugene~?6將質(zhì)粒瞬時(shí)轉(zhuǎn)染到293T細(xì)胞中,使用Western blotting和免疫熒光方法檢測(cè)目的基因的表達(dá)。結(jié)果發(fā)現(xiàn)克隆得到的梅花鹿Tβ10基因長(zhǎng)度為129bp,編碼42個(gè)氨基酸,成功構(gòu)建真核表達(dá)載體VR1012-Tβ10-HA,轉(zhuǎn)染后使用Western blotting方法檢測(cè)到梅花鹿Tβ10在293T細(xì)胞中表達(dá),免疫熒光方法證明梅花鹿Tβ10主要定位在細(xì)胞漿中。
[Abstract]:In order to study the relationship between T 尾 10 and the growth and development of velvet antler, the eukaryotic expression vector of T 尾 10 was constructed. The T 尾 10 gene of sika deer was amplified by Trizol. The target fragment was inserted into eukaryotic expression vector VR1012 to construct the expression plasmid. The plasmid was transiently transfected into 293T cells by Fugene~?6. Western blotting and immunofluorescence were used to detect the expression of the target gene. The results showed that the length of T 尾 10 gene of sika deer was 129 BP, encoding 42 amino acids, and the eukaryotic expression vector VR1012-T 尾 10 HA was successfully constructed. After transfection, the expression of T 尾 10 in 293T cells was detected by Western blotting. Immunofluorescence assay showed that T 尾 10 was mainly localized in the cytoplasm of sika deer.
【作者單位】： 長(zhǎng)春中醫(yī)藥大學(xué)中醫(yī)藥與生物工程研究開發(fā)中心;
【基金】：吉林省科技發(fā)展計(jì)劃(20140622003JC)
【分類號(hào)】：Q78;S825
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本文編號(hào)：1818567