本文選題：豬　切入點(diǎn)：FAMB基因　出處：《華南農(nóng)業(yè)大學(xué)學(xué)報(bào)》2017年03期 　論文類型：期刊論文

【摘要】：【目的】獲得豬FAM213B基因完整mRNA和啟動(dòng)子序列,研究豬FAM213B基因表達(dá),為探討母豬妊娠的建立和胚胎發(fā)育調(diào)控機(jī)制奠定基礎(chǔ)。【方法】通過5'RACE和3'RACE技術(shù),獲得基因完整mRNA序列,分析不同物種該基因氨基酸序列相似性;通過PCR克隆啟動(dòng)子區(qū),并通過雙熒光素酶報(bào)告基因載體系統(tǒng)轉(zhuǎn)染豬子宮內(nèi)膜細(xì)胞,研究其轉(zhuǎn)錄活性�！窘Y(jié)果】豬FAM213B基因mRNA全長(zhǎng)808 bp,其中5'UTR、CDS區(qū)和3'UTR長(zhǎng)度分別為67、609(含終止密碼子)和132 bp(不含poly A序列),在17~106位氨基酸之間存在硫氧還蛋白折疊結(jié)構(gòu)域;與豬FAM213B基因其他2個(gè)潛在轉(zhuǎn)錄本相比,三者都包含硫氧還蛋白折疊結(jié)構(gòu)域,但蛋白三級(jí)結(jié)構(gòu)存在較大差異;豬FAM213B氨基酸序列與山羊、牛和綿羊高度相似,相似性分別為94.03%、93.03%和91.54%。克隆獲得2 261 bp(-2 231/+30)的基因啟動(dòng)子序列,將其連接至雙熒光素酶報(bào)告基因載體,轉(zhuǎn)染豬子宮內(nèi)膜細(xì)胞,發(fā)現(xiàn)獲得的啟動(dòng)子片段能夠啟動(dòng)下游報(bào)告基因的轉(zhuǎn)錄,在啟動(dòng)子區(qū)存在潛在的典型NFκB等轉(zhuǎn)錄因子結(jié)合位點(diǎn)。【結(jié)論】本研究獲得豬FAM213B基因轉(zhuǎn)錄本長(zhǎng)度為808 bp,其蛋白存在硫氧還蛋白折疊功能結(jié)構(gòu)域,其啟動(dòng)子序列(-2 231/+30)在豬子宮內(nèi)膜細(xì)胞中具有較強(qiáng)的轉(zhuǎn)錄活性。
[Abstract]:[objective] to obtain the complete mRNA and promoter sequence of porcine FAM213B gene and study the expression of porcine FAM213B gene in order to lay a foundation for the establishment of pregnancy and the regulation mechanism of embryo development in sows. [methods] the complete mRNA sequence was obtained by 5race and 3RACE techniques. The amino acid sequences of different species were analyzed, the promoter region was cloned by PCR, and the porcine endometrial cells were transfected by double luciferase reporter gene vector system. [results] the total length of porcine FAM213B gene mRNA was 808 BP, in which the length of 5 UTRT CDS region and 3 FAM213B UTR region were 67 / 609 (including termination codon) and 132bp (excluding poly A) respectively, and there was a thioredoxin protein folding domain between the amino acids of 1710 ~ 6 amino acids. Compared with the other two potential transcripts of porcine FAM213B gene, all of them contain thioredoxin folding domain, but there is great difference in the tertiary structure of protein, and the amino acid sequence of porcine FAM213B is highly similar to that of goat, cattle and sheep. The similarity was 94.03% and 91.54%, respectively. The promoter sequence of 2 261 bp(-2 231 / 30) was cloned and ligated into double luciferase reporter gene vector and transfected into porcine endometrial cells. The obtained promoter fragment was found to be able to activate transcription of the downstream reporter gene. There are potential typical NF 魏 B transcription factor binding sites in the promoter region. [conclusion] the length of porcine FAM213B gene transcripts is 808 BP, and the protein has thioredoxin folding functional domain. The promoter sequence of the promoter-2 231 / 30) has a strong transcriptional activity in porcine endometrial cells.
【作者單位】： 廣東第二師范學(xué)院生物與食品工程學(xué)院/廣東高校應(yīng)用生態(tài)工程技術(shù)開發(fā)中心;華南農(nóng)業(yè)大學(xué)動(dòng)物科學(xué)學(xué)院/廣東省農(nóng)業(yè)動(dòng)物基因組學(xué)與分子育種重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金(31201771) 現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系專項(xiàng)(CARS-36)
【分類號(hào)】：Q78;S828

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