Localization of specific mRNAs in Xenopus embryos by whole-mount in situ hybridization.
Localization of specific mRNAs in Xenopus embryos by whole-mount in situ hybridization.
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发表时间：
1990-10
期刊：
影响因子：
4.6
通讯作者：
A. Hemmati-Brivanlou;D. Frank;M. E. Bolce;Bob D. Brown;Hazel;L. Sive;R. Harland
中科院分区：
文献类型：
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作者：
A. Hemmati-Brivanlou;D. Frank;M. E. Bolce;Bob D. Brown;Hazel;L. Sive;R. Harland
我们采用了一种非放射性技术来检测整个非洲爪哇胚胎中的定位mRNAs。在地高辛-UTP存在下转录的合成反义RNA被用作探针，并通过抗地高辛抗体进行检测。我们发现，从原肠发育晚期到蝌蚪阶段，可以检测到局部的mRNAs，并且可以检测到高丰度和低丰度的RNAs。该方法在肌肉肌动蛋白和α-珠蛋白RNA上进行了测试，它们的定位以前已经被描述过。此外，我们还利用该方法测定了XA-1RNA的分布，XA-1RNA是一种前外胚层特异的RNA，我们发现它在水泥腺的外围和孵化腺的区域都有表达。XA-1基因的序列预测了一种富含脯氨酸和组氨酸的蛋白质。
We have adapted a non-radioactive technique to detect localized mRNAs in whole-mount Xenopus embryos. Synthetic antisense RNA transcribed in the presence of digoxygenin-UTP is used as a probe and is detected via an anti-digoxygenin antibody. We show that localized mRNAs can be detected from late gastrula to tadpole stages and that high as well as low abundance RNAs can be detected. The method was tested on muscle actin and alpha-globin RNAs, whose localization has previously been characterized. In addition, we used the method to determine the distribution of XA-1 RNA, an anterior ectoderm-specific RNA, which we show is expressed in the periphery of the cement gland as well as in the region of the hatching gland. The sequence of an XA-1 cDNA predicts a protein rich in proline and histidine.