Isolation of the human gene that complements a temperature-sensitive cell cycle mutation in BHK cells.

M Ittmann; A Greco; C Basilico

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Isolation of the human gene that complements a temperature-sensitive cell cycle mutation in BHK cells.

机译：单独分离人类基因，其补充BHK细胞中的温度敏感细胞周期突变。

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We have cloned the human genomic DNA and the corresponding cDNA for the gene which complements the mutation of tsBN51, a temperature-sensitive (Ts) cell cycle mutant of BHK cells which is blocked in G1 at the nonpermissive temperature. After transfecting human DNA into TsBN51 cells and selecting for growth at 39.5 degrees C, Ts+ transformants were identified by their content of human AluI repetitive DNA sequences. Following two additional rounds of transfection, a genomic library was constructed from a tertiary Ts+ transformant and a recombinant phage containing the complementing gene isolated by screening for human AluI sequences. A genomic probe from this clone recognized a 2-kilobase mRNA in human and tertiary transformant cell lines, and this probe was used to isolate a biologically active cDNA from the Okayama-Berg cDNA expression library. Sequencing of this cDNA revealed a single open reading frame encoding a polypeptide of 395 amino acids. The deduced BN51 gene product has a high proportion of acidic and basic amino acids which are clustered in four hydrophilic domains spaced at 60- to 80-amino-acid intervals. These domains have strong sequence homology to each other. Thus, the tsBN51 protein consists of periodic repetitive clusters of acidic and basic amino acids.

机译：我们已经克隆了人类基因组DNA和对基因的相应cDNA，其补充了TSBN51的突变，在非智能温度下在G1中封闭的BHK细胞的温度敏感（TS）细胞周期突变体。将人DNA转染到TSBN51细胞中并选择39.5℃的生长后，通过它们的人alui重复DNA序列的含量鉴定TS +转化体。在另外两轮转染后，基因组文库由叔Ts +转化体构成和含有通过筛选人氧化序列分离的补充基因的重组噬菌体。来自该克隆的基因组探针在人和叔转化细胞系中识别出2千碱基mRNA，并且该探针用于分离来自冈山-Berg cDNA表达文库的生物活性cDNA。该cDNA的测序揭示了编码395个氨基酸多肽的单个开放阅读框。推导的BN51基因产物具有高比例的酸性和碱性氨基酸，其在以60-至80-氨基酸间隔间隔开的四个亲水结构域中聚集。这些域彼此具有强序列同源性。因此，TSBN51蛋白由周期性重复簇组成的酸性和碱性氨基酸。

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《Molecular and Cellular Biology》 |1987年第10期|共8页
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M Ittmann; A Greco; C Basilico;
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1. Isolation of the human gene that complements a temperature-sensitive cell cycle mutation in BHK cells. [J] . M Ittmann, A Greco, C Basilico Molecular and Cellular Biology . 1987,第10期

机译：单独分离人类基因，其补充BHK细胞中的温度敏感细胞周期突变。
2. Isolation of the human gene that complements a temperature-sensitive cell cycle mutation in BHK cells. [J] . M Ittmann, A Greco, C Basilico Molecular and Cellular Biology . 1987,第10期

机译：单独分离人类基因，其补充BHK细胞中的温度敏感细胞周期突变。
3. Isolation of temperature-sensitive mutations in the c-raf-1 catalytic domain and expression of conditionally active and dominant-defective forms of Raf-1 in cultured mammalian cells. [J] . Lu KK, Bazarov AV, Yoon LS, Cell Growth & Differentiation: The Molecular Biology Journal of the American Association for Cancer Research . 1998,第5期

机译：在培养的哺乳动物细胞中，分离c-raf-1催化域中的温度敏感突变，并表达Raf-1的条件活性和显性缺陷形式。
4. The effects benzo(a)pyrenediolexpoxide exposure on DNA adduct formation, altered gene expression, and toxicity in human lymphoblastoid TK6 cells. [C] . Joshua J Klaene, Jennifer Barrett, Caroline Ceailles Flakaros, American Society for Mass Spectrometry Conference on Mass Spectrometry and Allied Topics . 2011

机译：苯并（a）Pyrenyiolexpatiene暴露在DNA加合物形成，改变基因表达和人淋巴细胞Tk6细胞中的效果暴露。
5. Isolation of a humancDNA for threonyl-tRNA synthetase: Amplification and regulation of the structural gene in Chinese hamster ovary cells. [D] . Kontis, Kris John. 1989

机译：苏糖基-tRNA合成酶的人类cDNA的分离：中国仓鼠卵巢细胞结构基因的扩增和调控。
6. Isolation of the human gene that complements a temperature-sensitive cell cycle mutation in BHK cells. [O] . M Ittmann, A Greco, C Basilico 1987

机译：分离与BHK细胞中温度敏感的细胞周期突变互补的人类基因。
7. Isolation of the human gene that complements a temperature-sensitive cell cycle mutation in BHK cells. [O] . Ittmann, M, Greco, A, Basilico, C 1987

机译：分离与BHK细胞中温度敏感的细胞周期突变互补的人类基因。
8. Isolation of Estrogen-Responsive Genes in Human Breast Cancer Cells. [R] . Davie, J. R. 2001

机译：人乳腺癌细胞中雌激素反应基因的分离。

Isolation of the human gene that complements a temperature-sensitive cell cycle mutation in BHK cells.

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