Multiple forms of human dihydrofolate reductase messenger RNA. Cloning and expression in Escherichia coli of their DNA coding sequence.
Multiple forms of human dihydrofolate reductase messenger RNA. Cloning and expression in Escherichia coli of their DNA coding sequence.
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多种形式的人二氢叶酸还原酶信使 RNA。
DOI：
10.1016/0022-2836(82)90268-6
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发表时间：
1982
影响因子：
5.6
通讯作者：
Attardi,G
中科院分区：
文献类型：
--
作者：
Morandi,C;Masters,JN;Mottes,M;Attardi,G
已发现，与来自其亲本系（VA 2-B）的RNA相比，在来自甲氨蝶呤抗性人细胞变体（6A 3）的含多聚腺苷酸的RNA中，在兔网织红细胞裂解物中合成人二氢叶酸还原酶的编程能力大大增强。这种促进活性的一个主要部分与一个3.8 × 103碱基的RNA种类有关，在6A 3细胞的RNA的溴化乙锭染色电泳图中可检测到一条带，但在VA 2-B细胞的RNA中却没有。此外，蔗糖梯度分离实验表明，信使活性的另一个重要部分与大小约为103个碱基的RNA组分有关。从6A 3细胞的含poly（A）的总RNA合成的双链互补DNA已被大小分级，并且大尺寸（1400至3800碱基对）和小尺寸互补DNA（600至1400碱基对）种类已分别用于以pBR 322作为载体的大肠杆菌χ2282的转化。在用大尺寸互补DNA获得的76个转化体中，通过差异菌落杂交测定鉴定，没有一个转化体表达E.大肠杆菌，如判断耐药性甲氧苄啶。相比之下，8甲氧苄啶抗性转化体已获得使用小尺寸互补DNA，其质粒已被证明含有二氢叶酸还原酶编码序列的限制性作图和DNA测序，此外，免疫放射自显影实验揭示了存在于提取物中的两个这些转化体的蛋白质的电泳迁移率和免疫反应性的人二氢叶酸还原酶。限制性酶切图谱和DNA转移杂交实验进一步表明，赋予宿主甲氧苄氨嘧啶抗性的嵌合质粒和缺乏这种能力的嵌合质粒的插入片段共同覆盖了约3.35 × 103 bp的互补DNA区域，其中564 bp的二氢叶酸还原酶编码段位于正义链的5′端附近。使用不同克隆的互补DNA片段作为探针的RNA转移杂交实验表明，在来自两种甲氨蝶呤耐药变体6A 3和10 B3的含poly（A）的RNA中，存在三种二氢叶酸还原酶特异性信使RNA，大小分别为3.8 × 103、1.0 × 103和0.8 × 103个碱基，3 '非翻译区的长度不同，并且，在来自它们各自的亲本VA 2B和HeLa BU 25的RNA中的量大大减少。
The programming capacity for the synthesis of human dihydrofolic acid reductase in a rabbit reticulocyte lysate has been found to be greatly enhanced in the polysomal poly(A)-containing RNA from a methotrexate-resistant human cell variant (6A3), as compared to the RNA from its parental line (VA2-B). A major fraction of this promoting activity is associated with a 3.8 × 103base RNA species detectable as a band in the ethidium bromide-stained electrophoretic pattern of the RNA from 6A3 cells, but not in the RNA from VA2-B cells. Furthermore, sucrose gradient fractionation experiments have indicated that another substantial portion of the messenger activity is associated with RNA components around 103bases in size. Double-stranded complementary DNA synthesized from total poly(A)-containing RNA of 6A3 cells has been size fractionated, and both large (1400 to 3800 base-pairs) and small size complementary DNA (600 to 1400 base-pairs) species have been used separately to transformEscherichia coliχ2282 with pBR322 as a vector. Of 76 transformants obtained with the large size complementary DNA, identified by a differential colony hybridization assay, none has expressed the dihydrofolic acid reductase coding sequence inE. coli, as judged by resistance to trimethoprim. By contrast, eight trimethoprim-resistant transformants have been obtained using the small size complementary DNA, and their plasmids have been shown to contain the dihydrofolic acid reductase coding sequence by restriction mapping and DNA sequencing; moreover, immunoautoradiographic experiments have revealed the presence in the extracts of two of these transformants of a protein with the electrophoretic mobility and immunoreactivity of human dihydrofolic acid reductase. Restriction mapping and DNA transfer hybridization experiments have further indicated that the inserts of the chimaeric plasmids conferring trimethoprim resistance upon the host and of those lacking this capacity cover together a complementary DNA region of about 3.35 × 103base-pairs, in which the 564 base-pair dihydrofolic acid reductase coding stretch is located near the 5′ end of the sense strand. RNA transfer hybridization experiments using different cloned complementary DNA fragments as probes have shown the presence of three species of dihydrofolic acid reductase-specific messenger RNAs, with sizes of 3.8 × 103, 1.0 × 103and 0.8 × 103bases, differing in the length of the 3′ untranslated region, in the poly(A)-containing RNA from two methotrexate-resistant variants, 6A3 and 10B3, and, in greatly reduced amounts, in the RNA from their respective parents, VA2B and HeLa BU25.
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小鼠淋巴瘤 L1210 的二氢叶酸还原酶的氨基酸序列。
DOI：
10.1016/s0021-9258(17)37942-5
发表时间：
1979
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
D. Stone;S. Paterson;J. H. Raper;A. W. Phillips
通讯作者：
A. W. Phillips
影响因子：
5.6
作者：
AMALRIC, F;MERKEL, C;ATTARDI, G
通讯作者：
ATTARDI, G
影响因子：
4.4
作者：
S. Kessler
通讯作者：
S. Kessler
β-珠蛋白 mRNA 的 3 非编码区对于体外翻译不是必需的
影响因子：
14.9
作者：
M. Kronenberg;B. Roberts;A. Efstratiadis
通讯作者：
A. Efstratiadis
用 SV40 转化的人体组织培养物的形态学和病毒学研究。
影响因子：
--
作者：
J. Pontén;F. Jensen;H. Koprowski
通讯作者：
H. Koprowski