大鼠胆碱酯酶mRNA在卵母细胞中的表达

EXPRESSION OF ACTIVE ACETYLCHOLINESTERASE IN XENOPUS OOCYTESMICROINJECTED WITHmRNA FROM RAT BRAIN

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。

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).