Affinity chromatography—dependent selection （ACDS） of genomic DNA fragments bound specifically to bacterial synthesized Myc／Myn proteins

This paper describes an approach to seek for mouse c-Myc/Myn proteins-bound specific sequences among genomic DNA.cDNA fragment of myn gene was obtained through RT-PCR technique from RNA of NIH3T3 cells.DNA fragments encoding BR/HLH/LZ structure of Myc and Myn proteins were cloned in frame into pGEX-2T vector respectively.Fusion GST-Myc and GST-Myn synthesized in E.coli hosts showed affinity to CACGTG E-box DNA and subsequently interacted with genomic fragments prepared through whole-genome-PCR.A PCR-assisted procedure which combines protein-DNA interaction and affinity chromatography was designed to enrich Myc/Myn bound DNA.At least two genomic DNA fragments obtained exhibit specifical binding capacity to Myc/Myn complex but not to GST alone.Significance of the work and of the technique itself as well asidentification of the DNAs are discussed.     

细叶黄芪叶肉原生质体发育早期几种细胞器的变化

在细叶黄芪叶肉原生质体发育早期,细胞器的变化较大。离体培养4h后,线粒体的嵴和基质物质开始增加。培养3—5天后,线粒体的数量增加5倍以上,此时可见大部分线粒体围绕细胞核分布。在培养24h后,高尔基体开始发育,它们主要分布在细胞质周边区域。多糖细胞化学染色表明,高尔基体内沉积着大量嗜银物质。培养1天后,粗面内质网开始发育。培养3天时,部分叶绿体边缘出现一些空隙结构。随着叶绿体内膜结构的消失,淀粉粒增大,叶绿体逐渐转变为造粉质体。     

The inhibitory effect of transthyretin gene on growth of human hepatoma cells

Transthyretin(TTR) gene was highly expressed in normal liver and it has been found to be deleted in part of DNA samples from human hepatic cancer.Its mRNA expression was suppressed in most hepatoma samples.In order to study the biological effect of TTR gene on the growth of hepatoma cells,a recombinant vector containing TTR cDNA was constructed by pCMV,then it was transfected into hepatoma cell lines SMMC-7721 and Q3.It has been demonstrated that the inhibition of growth rate of TTR cDNA transfected hepatoma cells was about 50% in strength compared with that of the control.This inhibition was further enhanced when the transfected hepatoma cells were treated with all-trans retinoic acid.Hepatoma cells of cell lines PLC/PRF/5,SMMC-7721 and Q3 as well as hepatoma cells SMMC-7721 transfected with pCMV or pCMV-TTR were analyzed for TTR expression by Northern hybridization.The low level of TTR expression was found in both hepatoma cell lines and in SMMC-7721 cells transfected with pCMV alone.However,a remarkable TTR mRNA expression was observed in hepatoma SMMV-7721 cells transfected with pCMV-TTR.It seems possible that TTR gene might be a candidate of cancer suppressor gene for human hepatic cancer.     

A stage—specific protein factor binding to a CACCC motif in both human β—globin gene promoter and 5‘—HS2 region

The DNaseI hypersensitive site 2 (HS2) of human β-globin locus control region(LCR) is required for the high level expression of human β-globin genes.In the present study,a stage-specific protein factor (LPF-β) was identified in the nuclear extract prepared from mouse fetal liver at d 18 of gestation,which could bind to the HS2 region of human β-globin LCR.We also found that the shift band of LPF-β factor could be competed by human β-globin promoter.However,it couldn‘t be competed by human ε-globin promoter or by human ^Aγ-globin promoter.Furthermore,our data demonstrated that the binding-sequence of LPF-β factor is 5‘CACACCCTA 3‘,which is located at the HS2 region of β-LCR(from-10845 to-10853 bp)and human β-globin promoter(from-92 to -84 bp).We speculated that these regions containing the CACCC box in both the human β-globin promoter and HS2 might function as stage selector elements in the regulation of human β-globin switching and the LPF-β factor might be a stage-specific protein factor involved in the regulation of human β-globin gene expression.     

在调查反馈的信息中探寻生物教学方向

生物学是当代科学研究的热点和前沿,它与人类面临的“环境”、“人口”、“粮食”、“能源”四大危机具有密切关系。为了提高国民素质,在基础教育中普及生物知识是非常必要的,但由于追求升学考试等现实问题,使生物学科在中学理科中的位置面临很严重的问题(与发达     

笼养间蜂猴的繁殖

1989年至今,对15只（10,5）成年间蜂猴在人工饲养条件下的繁殖进行观察,结果为：1．间蜂猴的繁殖有明显的季节性;2．发情周期为49．67d(SD=1.25),在此期间,雌性外生殖器红肿,变大;雄性阴囊胀大;3．交配以背腹相贴为主;4．怀孕期为188d;5．哺乳期为108d(SD=4.12);6．均为一胎二仔。     

用藻类评价铬渣污染水体的恢复程度

受铬渣渗水污染17年的3个鱼塘,至今藻类群落结构已多样化发展,除了绿藻门(Chlorophyta)为优势类群,占所有藻类属数61.1%外,还有硅藻门(Bacillarlophyta)占16.7%,蓝藻门(Cyanophyta)占11%,隐藻门(Crytophyta)和裸藻(Euglenophyta)各占5.6%。3个鱼塘藻类群落中的 Shannon—Weaver 多样性指数,与水体铬污染大致成负相关关系.S—W 多性指效,虽然冬、夏有差异,但冬天2号塘可升至3左右鉴于铬渣渗水污染的3个鱼塘,经过17年以后铬元素浓度已显著降低,量分别为0.0044mg/L,0.0056mg/L 和0.0069mg/L,已降至允许的范围,因此,作者认为该地区铬渣严重污染的水体有明显(?),已恢复至轻度污染的程度.