發(fā)布時間：2018-04-29 05:18

  本文選題：梅花鹿 + 胸腺素beta　； 參考：《生物技術通報》2017年06期

【摘要】：轉錄組測序發(fā)現(xiàn)Tβ10在鹿茸中高表達,為了研究Tβ10與鹿茸生長發(fā)育間的關系,構建Tβ10真核表達載體。使用Trizol法提取梅花鹿鹿茸總RNA,PCR技術特異性擴增梅花鹿Tβ10基因,利用酶切位點將目的片段插入真核表達載體VR1012構建表達質粒,通過Fugene~?6將質粒瞬時轉染到293T細胞中,使用Western blotting和免疫熒光方法檢測目的基因的表達。結果發(fā)現(xiàn)克隆得到的梅花鹿Tβ10基因長度為129bp,編碼42個氨基酸,成功構建真核表達載體VR1012-Tβ10-HA,轉染后使用Western blotting方法檢測到梅花鹿Tβ10在293T細胞中表達,免疫熒光方法證明梅花鹿Tβ10主要定位在細胞漿中。
[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】： 長春中醫(yī)藥大學中醫(yī)藥與生物工程研究開發(fā)中心;
【基金】：吉林省科技發(fā)展計劃(20140622003JC)
【分類號】：Q78;S825
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本文編號：1818567