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孕期人胎盘催乳素mRNA及其结构基因:用互补DNA进行定量分析

Human placental lactogen mRNA and its structural genes during pregnancy: quantitation with a complementary DNA.

作者信息

McWilliams D, Callahan R C, Boime I

出版信息

Proc Natl Acad Sci U S A. 1977 Mar;74(3):1024-7. doi: 10.1073/pnas.74.3.1024.

Abstract

A complementary DNA (cDNA) strand was transcribed from human placental lactogen (hPL) mRNA. Based on alkaline sucrose gradient centrifugation, the size of the cDNA was about 8 S, which would represent at least 80% of the hPL mRNA. Previously we showed that four to five times more hPL was synthesized in cell-free extracts derived from term as compared to first trimester placentas. Hybridization of the cDNA with RNA derived from placental tissue revealed that there was about four times more hPL mRNA sequences in total RNA from term placenta than in a comparable quantity of total first trimester RNA. Only background hybridization was observed when the cDNA was incubated with RNA prepared from human kidney. To test if this differential accumulation of hPL mRNA was the result of an amplification of hPL genes, we hybridized the labeled cDNA with cellular DNA from first trimester and term placentas and with DNA isolated from human brain. In all cases, the amount of hPL sequences was approximately two copies per haploid genome. Thus, the enhanced synthesis of hPL mRNA appears to result from a transcriptional activation rather than an amplification of the hPL gene. The increase likely reflects placental differentiation in which the proportion of syncytial trophoblast increases at term.

摘要

从人胎盘催乳素(hPL)mRNA转录出一条互补DNA(cDNA)链。基于碱性蔗糖梯度离心法,cDNA的大小约为8S,这至少占hPL mRNA的80%。此前我们发现,与孕早期胎盘来源的无细胞提取物相比,足月胎盘来源的无细胞提取物中合成的hPL多四到五倍。cDNA与胎盘组织来源的RNA杂交显示,足月胎盘总RNA中的hPL mRNA序列比等量的孕早期总RNA中的多约四倍。当cDNA与从人肾制备的RNA孵育时,仅观察到背景杂交。为了测试hPL mRNA这种差异积累是否是hPL基因扩增的结果,我们将标记的cDNA与孕早期和足月胎盘的细胞DNA以及从人脑中分离的DNA进行杂交。在所有情况下,hPL序列的量约为每个单倍体基因组两份。因此,hPL mRNA合成的增强似乎是转录激活的结果,而不是hPL基因的扩增。这种增加可能反映了胎盘分化,足月时合体滋养层的比例增加。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eef/430573/bf367d7d70ec/pnas00025-0231-a.jpg

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