KLF7b对斑马鱼胚胎发育的影响
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江苏省高等学校自然科学研究重大项目(17KJA310001);江苏高校优势学科建设工程资助项目


Effects of KLF7b on embryo development of zebrafish
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    摘要:

    目的:探讨KLF7b对斑马鱼胚胎发育的影响。方法:体外转录野生型KLF7b mRNA、KLF7b 乙酰化位点突变体mRNA;设计合成特异性KLF7b反义吗啉代寡核苷酸(morpholino oligonueleitide,MO);在1~2细胞期注射到斑马鱼胚胎体内以实现KLF7b的过表达、位点突变与敲降;观察胚胎表型并利用定量PCR检测发育相关基因的表达变化,钙黄绿素染色观察斑马鱼骨骼发育情况。结果:过表达KLF7b对斑马鱼胚胎的发育无明显影响;敲降KLF7b或注射乙酰化位点突变的KLF7b后,斑马鱼胚胎出现心包积液、体轴和头部发育的异常,肌肉发育标志基因表达下调。结论:KLF7b可通过乙酰化修饰影响斑马鱼的发育。

    Abstract:

    Objective:To explore the effects of KLF7b on embryo development of zebrafish. Methods:Wild-type KLF7b mRNA and KLF7b acetylated site mutant mRNA were transcripted in vitro. Specific mRNA and morpholino oligonueleitides(MOs)were used to overexpress or site mutate or knockdown KLF7b by microinjection at 1~2 cells stage. The phenotypes of embryos were observed at different stages and the expressions of marker genes were tested by quantitative RT-PCR. The bone development of zebrafish was detected by calcein staining. Results:KLF7b overexpression had no effects on embryo development of zebrafish. KLF7b knockdown or acetylated site mutation caused embryonic abnormalities including pericardial effusion,the developmental malformations of somite and head. The marker genes of muscle were down-regulated. Conclusion:KLF7b may affect the development of zebrafish via its own acetylation.

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李德利,黄梦圆,魏丽圆,王 凯,王学军,陈园园,袁 栎. KLF7b对斑马鱼胚胎发育的影响[J].南京医科大学学报(自然科学版),2018,(5):605-611

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  • 收稿日期:2018-01-03
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  • 在线发布日期: 2018-05-16
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