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人血小板生成素的内含子V而非内含子I，可提高由山羊β-酪蛋白启动子调控的转基因小鼠乳汁中的表达。

Intron V, not intron I of human thrombopoietin, improves expression in the milk of transgenic mice regulated by goat beta-casein promoter.

作者信息

Li Yan, Hao Hu, Zhou Mingqian, Zhou Hongwei, Ye Jianbin, Ning Lijun, Ning Yunshan

机构信息

School of Biotechnology, Southern Medical University, North1838 Guangzhou Road, Guangzhou, 510515, PR. China.

Department of Pediatrics, the Six Affiliated Hospital of Sun Yatsen University, 26 Yun Cun Erheng Road, Guangzhou, 510655, PR. China.

出版信息

Sci Rep. 2015 Nov 3;5:16051. doi: 10.1038/srep16051.

DOI:10.1038/srep16051
PMID:26527459
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4630608/
Abstract

Introns near 5' end of genes generally enhance gene expression because of an enhancer /a promoter within their sequence or as intron-mediated enhancement. Surprisingly, our previous experiments found that the vector containing the last intron (intron V) of human thromobopoietin (hTPO) expressed higher hTPO in cos-1 cell than the vector containing intron I regulated by cytomegalovirus promoter. Moreover, regulated by 1.0 kb rat whey acidic protein promoter, hTPO expression was higher in transgenic mice generated by intron V-TPOcDNA than in transgenic mice generated by TPOcDNA and TPOgDNA. However, it is unknown whether the enhancement of hTPO expression by intron I is decreased by uAUG7 at 5'-UTR of hTPO in vivo. Currently, we constructed vectors regulated by stronger 6.5 kb β-casein promoter, including pTPOGA (containing TPOcDNA), pTPOGB (containing TUR-TPOcDNA, TUR including exon1, intron I and non-coding exon2 of hTPO gene), pTPOGC (containing ΔTUR-TPOcDNA, nucleotides of TUR from uAUG7 to physiological AUG were deleted), pTPOGD (containing intron V-TPOcDNA) and pTPOGE (containing TPOgDNA), to evaluate the effect of intron I on hTPO expression and to further verify whether intron V enhances hTPO expression in the milk of transgenic mice. The results demonstrated that intron V, not intron I improved hTPO expression.

摘要

基因5'端附近的内含子通常会增强基因表达，这是由于其序列中的增强子/启动子，或是通过内含子介导的增强作用。令人惊讶的是，我们之前的实验发现，含有人类血小板生成素（hTPO）最后一个内含子（内含子V）的载体在cos-1细胞中表达的hTPO比含有由巨细胞病毒启动子调控的内含子I的载体更高。此外，在1.0 kb大鼠乳清酸性蛋白酸性蛋白启动子的调控下，由内含子V-TPOcDNA产生的转基因小鼠中hTPO的表达高于由TPOcDNA和TPOgDNA产生的转基因小鼠。然而，在体内hTPO的5'-UTR处的uAUG7是否会降低内含子I对hTPO表达的增强作用尚不清楚。目前，我们构建了由更强的6.5 kbβ-酪蛋白启动子调控的载体，包括pTPOGA（含有TPOcDNA）、pTPOGB（含有TUR-TPOcDNA，TUR包括hTPO基因的外显子1、内含子I和非编码外显子2）、pTPOGC（含有ΔTUR-TPOcDNA，TUR从uAUG7到生理性AUG的核苷酸被删除）、pTPOGD（含有内含子V-TPOcDNA）和pTPOGE（含有TPOgDNA），以评估内含子I对hTPO表达的影响，并进一步验证内含子V是否能增强转基因小鼠乳汁中hTPO的表达。结果表明，是内含子V而非内含子I提高了hTPO的表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/419ba8b51f9a/srep16051-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/4b58d5c13f74/srep16051-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/c035185693a3/srep16051-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/a54bb609c667/srep16051-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/30486257068d/srep16051-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/419ba8b51f9a/srep16051-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/4b58d5c13f74/srep16051-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/c035185693a3/srep16051-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/a54bb609c667/srep16051-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/30486257068d/srep16051-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/466a/4630608/419ba8b51f9a/srep16051-f5.jpg

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本文引用的文献

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Production of recombinant human proinsulin in the milk of transgenic mice.在转基因小鼠的乳汁中生产重组人胰岛素原。
Sci Rep. 2014 Sep 30;4:6465. doi: 10.1038/srep06465.
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The last intron of the human thrombopoietin gene enhances expression in milk of transgenic mice.人血小板生成素基因的最后一个内含子可增强转基因小鼠乳汁中的表达。
Funct Integr Genomics. 2014 Mar;14(1):229-36. doi: 10.1007/s10142-013-0348-x. Epub 2013 Nov 28.
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The biology of thrombopoietin and thrombopoietin receptor agonists.血小板生成素及其受体激动剂的生物学特性。
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Give me an intron: any intron.给我一个内含子：任何内含子。
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Expression of the human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene under control of the 5'-regulatory sequence of the goat alpha-S1-casein gene with and without a MAR element in transgenic mice.在转基因小鼠中，人粒细胞-巨噬细胞集落刺激因子（hGM-CSF）基因在山羊α-S1-酪蛋白基因的 5'调控序列控制下，有无 MAR 元件的表达。
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The high-level expression of human tissue plasminogen activator in the milk of transgenic mice with hybrid gene locus strategy.利用杂种基因座策略在转基因小鼠的乳汁中高水平表达人组织型纤溶酶原激活物。
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