大鼠胆碱酯酶mRNA在卵母细胞中的表达

ＡＧＰＣ法是一种快速、简便、有效的ＲＮＡ提取方法。参照该法自新生大鼠脑组织中提取出总ＲＮＡ，进一步通过ｏｌｉｇｏ（ｄＴ）－纤维素亲和层析分离获得ｐｏｌｙ（Ａ）￣＋ＲＮＡ。当ｐｏｌｙ（Ａ）￣＋ＲＮＡ或总ＲＮＡ经显微注射法进入非洲爪蟾卵母细胞后，其翻译系统能够利用外源ｍＲＮＡ合成具有催化活性的胆碱酯酶。表达的胆碱酯酶大部分分泌到卵母细胞培养液中。在一定范围内，ｍＲＮＡ的注射量与胆碱酯酶的表达量成正比。所合成的胆碱酯酶活性可被毒扁豆碱和Ｓ－（２－二异丙基氨乙基）甲基硫赶膦酸乙酯（ＶＸ）抑制。

EXPRESSION OF ACTIVE ACETYLCHOLINESTERASE IN XENOPUS OOCYTESMICROINJECTED WITHmRNA FROM RAT BRAIN

Using AGPC method,a rapid,simple and efficient way of RNA isolation,total RNA was extracted from neonatal rat brain and poly(A) RNA was further purifled using oligo dT cellulose affinity chromatography.When Xenopus oocytes were microinjected with RNA from rat brain,they synthesized catalytically active acetylcholinesterase.The synthesized enzyme could be se-creted into oocvte culture medium.There was a dependence of the enzvme expression on the amount of injected RNA.The enzyme activitv could be inhibited by eserine and S-2-diisopropylaminoethyl methyl phosphonothiolate(VX).