摘要
自２例入脑胶质瘤手术标本中纯化ｍＲＮＡ，逆转录合成双链ｃＤＮＡ分子。在Ｔ４ＤＮＡ连接酶作用下，与Ａｄａｐｔｏｒ插头相连。经Ｔ４多聚核苷酸激酶磷酸化后，再与脱磷酸处理过的γｇｔ１１臂连接，得到重组γｇｔ１１ＤＮＡ。体外包装，构建了一个库容量含１．０２×１０＾６重组子的人脑胶质瘤ｃＤＮＡ基因文库，该文库的克隆效率为１．０８×１０７ｐｆｕ／μｇ ｃＤＮＡ，重组百分比为９６．２％。并初步筛选出了胶质纤维…
mRNA molecules were extracted fromtwo operative specimens of human glioma.The double-stranded cDNA molecules weresynthesized by reverse transcription.The cD-NA molecules were ligated to adaptors by T_4DNA ligase.After the adapted cDNAmolecules has been kinased,they could be lig-ated to dephosphorylated λgt11 vector arms.This resulted in recombinant λgt11 DNAmolecules.The human glioma cDNA libraryconstructed by in vitro packaging.The librarycontained 1.02×10~6 recombinants.Cloningefficiency was 1.08×10~7pfu/μgcDNA.Per-centage of recombinants was 96.2%.The li-brary contained the clones corresponded glialfibrillary acidic protein by initial immuno-screening.Its frequency was 0.02% on thelevel of expression.
出处
《中华神经外科杂志》
CSCD
北大核心
1993年第5期272-275,共4页
Chinese Journal of Neurosurgery
基金
国家自然科学基金
