人YAP1基因亚型的质粒构建及表达鉴定

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Abstract Objective: To construct two isoforms of YAP1 for further research into their difference. Methods: YAP1-1δ and YAP1-2δ fragments were obtained by using overlapping PCR. And then the two DNA fragments were respectively ligated to the plasmid contained partial YAP1 gene via ClaI and SalI two restriction enzyme cutting sites. The recombinant vectors named pCI2-Flag-YAP1-1δ and pCI2-Flag-YAP1-2δ were identified by restriction enzyme digestion and gene sequencing. After transfection of 293 cells with the two vectors for 48 h, the expression of YAP1 protein was detected by Western blot. Results: The two over-expressing plasmids were successfully constructed and YAP1-1δ and YAP1-2δ proteins were strongly expressed in HEK293 cells. Conclusion:
The HEK293 cells could transiently express YAP1-1δ and YAP1-2δ by using the recombinant plasmids pCI2-Flag-YAP1-1δ and pCI2-Flag-YAP1-2δ, which lays a foundation for further study of the different isoforms of YAP1 and its mechanism in the development of tumorigenesis.

Key words： YAP1 gene overlapping PCR eukaryotic expression recombinant vector HEK293 cells

Received: 04 December 2017

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	Articles by authors
	DONG Jinglai
	ZHANG Jinsan
	GUO Qiang
	CHEN Qianjie
	GUO Lisha
	LI Xiaokun.

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2018/V48/I4/241

[1] SANDESC M, DINU A, ROGOBETE A F, et al. Circulating microRNAs expressions as genetic biomarkers in pancreatic cancer patients continuous non-invasive monitoring[J]. Clin Lab, 2017, 63(10): 1561-1566.
[2] CHEN Y, SHI Y, YAN H, et al. Timing of chemotherapy-induced neutropenia: the prognostic factor in advanced pancreatic cancer patients treated with gemcitabine/gemcitabine-based chemotherapy[J]. Oncotarget, 2017, 8(39): 66593-66600.
[3] ZHANG W, NANDAKUMAR N, SHI Y, et al. Downstream of mutant KRAS, the transcription regulator YAP is essential for neoplastic progression to pancreatic ductal adenocar-cinoma[J]. Sci Signal, 2014, 7(324): ra42.
[4] NGUYEN L T, TRETIAKOVA M S, SILVIS M R, et al. ERG activates the YAP1 transcriptional program and induces the development of age-related prostate tumors[J]. Cancer Cell, 2015, 27(6): 797-808.
[5] MELFI S, COLCIAGO A, GIANNOTTI G, et al. Stressing out the Hippo/YAP signaling pathway: toward a new role in Schwann cells[J]. Cell Death Dis, 2015, 6: e1915.
[6] CAO X, PFAFF S L, GAGE F H. YAP regulates neural progenitor cell number via the TEA domain transcription factor [J]. Genes Dev, 2008, 22(23): 3320-3334.
[7] PLOUFFE S W, HONG A W, GUAN K L. Disease implications of the Hippo/YAP pathway[J]. Trends Mol Med, 2015, 21(4): 212-222.
[8] SUDOL M. YAP1 oncogene and its eight isoforms[J]. Oncogene, 2013, 32(33): 3922.
[9] SUDOL M. Newcomers to the WW domain-mediated network of the hippo tumor suppressor pathway[J]. Genes Cancer, 2010, 1(11): 1115-1118.
[10] 丁鑫, 孟刚, 童晓玲, 等. WW结构域与Hippo信号通路[J]. 蚕业科学, 2013, 39(6): 1177-1185.
[11] 李敏睿, 张盛洪, 钟碧慧. Hippo信号通路在恶性肿瘤及干细胞中的作用[J]. 胃肠病学和肝病学杂志, 2013, 22(4): 389-392.
[12] 邱勤明, 石秋燕, 王江, 等. Hippo信号通路与肿瘤发生的研究进展[J]. 四川生理科学杂志, 2016, 38(1): 35-38.
[13] HALL C A, WANG R, MIAO J, et al. Hippo pathway effector Yap is an ovarian cancer oncogene[J]. Cancer Res, 2010, 70(21): 8517-8525.
[14] 白帆, 刘小龙, 易俊, 等. Hippo通路与恶性肿瘤侵袭转移的相关性研究进展[J]. 癌症进展, 2016, 14(3): 213-216.
[15] 史梦婕, 邵松军, 李洁媚, 等. Hippo通路与肿瘤相关性研究进展[J]. 中国细胞生物学学报, 2014, 36(3): 361-365.
[16] 吴卉, 花荣, 陈锦先. Hippo通路及靶因子YAP与肿瘤关系的研究进展[J]. 实用医学杂志, 2013, 29(4): 515-517.
[17] 周广玺, 张翠萍, 魏文超, 等. 慢性胃炎及微环境与胃癌[J]. 齐鲁医学杂志, 2014, 29(2): 177-180.
[18] 王杰, 喻超, 周显飞, 等. MiRNA-138-5p抑制人胰腺癌细胞增殖能力[J]. 世界华人消化杂志, 2016, 24(28): 3970-3977.
[19] 周青峰, 罗慧, 朱雪琼. Yes相关蛋白在妇科恶性肿瘤中的研究进展[J]. 温州医科大学学报, 2016, 46(7): 543-547.
[20] PAN D. The hippo signaling pathway in development and cancer[J]. Dev Cell, 2010, 19(4): 491-505.
[21] LEUNG C Y, ZERNICKA-GOETZ M. Angiomotin prevents pluripotent lineage differentiation in mouse embryos via Hippo pathway-dependent and -independent mechanisms[J]. Nat Commun, 2013, 4: 2251.
[22] ARAGóN E, GOERNER N, ZAROMYTIDOU A I, et al. A Smad action turnover switch operated by WW domain readers of a phosphoserine code[J]. Genes Dev, 2011, 25(12): 1275-1288.
[23] LEHMANN W, MOSSMANN D, KLEEMANN J, et al. ZEB1 turns into a transcriptional activator by interacting with YAP1 in aggressive cancer types[J]. Nat Commun, 2016, 7: 10498.