本文選題：豬　切入點：ZBED基因　出處：《中國畜牧雜志》2017年08期 　論文類型：期刊論文

【摘要】：鋅指蛋白ZBED6基因的突變可促進骨骼肌中胰島素樣生長因子2(IGF2)轉錄水平顯著上調,從而促進細胞增殖和肌管形成。本研究利用CRISPR-Cas9技術對豬的ZBED6基因進行編輯,利用在線軟件篩選出4個突變靶點,利用p Cas9/g RNA載體構建p Cas9/g RNA-ZBED6質粒,再構建4個用于驗證突變效率的p TYNE-ZBED6質粒,同時轉染HEK293細胞系,通過觀察熒光信號的強弱判斷靶序列突變效率的高低。結果表明:位于編碼區(qū)第17~37位點的靶點1轉染后熒光信號最強,切割效率最高,該位點是利用CRISPR-Cas9技術編輯ZBED6基因的有效位點。
[Abstract]:The mutation of zinc finger protein ZBED6 gene can significantly up-regulate the transcription level of insulin-like growth factor-2 IGF2 in skeletal muscle, thus promote cell proliferation and myotube formation. In this study, CRISPR-Cas9 technique was used to edit porcine ZBED6 gene. Four mutation targets were screened out by on-line software, then p Cas9/g RNA-ZBED6 plasmid was constructed by using p Cas9/g RNA vector, and four p TYNE-ZBED6 plasmids were constructed to verify the mutation efficiency. At the same time, they were transfected into HEK293 cell line. The mutation efficiency of target sequence was judged by observing the intensity of fluorescence signal. The results showed that target 1, located at site 1737 of coding region, had the strongest fluorescence signal and the highest cutting efficiency after transfection. This site is an effective site for editing ZBED6 gene by CRISPR-Cas9 technique.
【作者單位】： 黑龍江省農(nóng)業(yè)科學院畜牧研究所;農(nóng)業(yè)部畜禽遺傳資源與種質創(chuàng)新重點實驗室;
【基金】：農(nóng)業(yè)部畜禽遺傳資源與種質創(chuàng)新重點實驗室開放課題 國家生豬產(chǎn)業(yè)技術體系(CARS-36)
【分類號】：Q78;S828

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1 楊野喬;基于嵌入式的熒光信號采集系統(tǒng)[D];電子科技大學;2011年

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