首页> 外文期刊>Journal of Biosciences >In vitro translation of RNA to lactase during postnatal development of rat intestine.
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In vitro translation of RNA to lactase during postnatal development of rat intestine.

机译:在大鼠肠的出生后将RNA体外翻译为乳糖酶。

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摘要

MRNA levels encoding lactase were detected by Northern blot analysis using two different probes in developing rat intestine. Probe I and probe II corresponding to second half of prolactase gene showed a 6.8 kb mRNA transcript in 7, 14, 21 and 30 day old rat intestine. There was no change in quantity of lactase mRNA detected using probe II, but hybridization with probe I showed a progressive decrease in mRNA transcript encoding lactase with age. At day 7 and 14 of postnatal development, the lactase mRNA was quite high, but it reduced upon weaning. The in vitro translation products of RNA detected by Western blot analysis using brush border lactase antibodies showed several isoforms of lactase antigen with molecular weight ranging from 100-220 kDa. Analysed at days 7 and 30 of postnatal development, lactase isoforms of molecular weight 130 kDa and 220 kDa were similar to those found in purified brush border membranes. The translation of RNA to 220 kDa lactase protein was high in 7 and 14 day old pups, but it was markedly reduced in 30 day old animals. These findings support the contention that translation of mRNA to lactase is impaired in weaned animals, which may also be responsible for the maturational decline in lactase activity in adult rat intestine.
机译:通过RNA印迹分析,使用发育中的大鼠肠中的两种不同探针,检测编码乳糖酶的MRNA水平。对应于乳糖酶基因后半部分的探针I和探针II在7、14、21和30天大的大鼠肠中显示6.8 kb mRNA转录本。使用探针II检测到的乳糖酶mRNA的数量没有变化,但是与探针I的杂交显示随着年龄的增长,编码乳糖酶的mRNA转录物逐渐减少。在产后发育的第7和14天,乳糖酶mRNA很高,但在断奶后会降低。使用刷状边缘乳糖酶抗体通过蛋白质印迹分析检测到的RNA的体外翻译产物显示了几种乳糖酶抗原的同工型,分子量范围为100-220 kDa。在产后发育的第7和30天进行分析,分子量130 kDa和220 kDa的乳糖酶同工型与纯化的刷状缘膜中发现的相似。在7和14日龄的幼崽中,RNA向220 kDa乳糖酶蛋白的翻译很高,但在30日龄的动物中,RNA的翻译明显减少。这些发现支持断奶动物中mRNA向乳糖酶的翻译受损的论点,这也可能是成年大鼠肠中乳糖酶活性的成熟下降的原因。

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