降血脂药物对酰基CoA氧化酶基因反式作用因子的影响

从对照和用ＤＥＨＰ处理的大鼠肝脏提取核蛋白,以含酰基ＣｏＡ氧化酶基因表达调控部位的ＤＮＡ片段和该基因的不同蛋白结合位点的ＤＮＡ片段作为核蛋白结合反应的探针,通过凝胶电过移率改变实验和Ｓｏｕｔｈｗｅｓｔｅｒｎ印迹分析检查了ＤＥＨＰ对ＡＯＸ基因反式作用因子的含量和（或）与基因的结合活性,在转录水平上促进基因的表达。     

大鼠C3基因TATA区结合核蛋白与其转录活性关系的研究

大鼠Ｃ３基因的转录受雄激素调控且只在大鼠腹侧前列腺中表达。本文报道用凝胶电泳带漂移分析和离体ＤＮａｇｅＩ足迹法分析研究了大鼠Ｃ３基因ＴＡＴＡ区结合核蛋白与其组织专一的和雄激素调控的转录活性间的关系。结果表示：相同的（－５０到－１０）４ｂｐ的ＴＡＴＡ区ＤＮＡ片段在不同组织的细胞核中结合的核蛋白是不同的。通过与一个仅在ＴＡＴＡ区－２９．位核苷酸Ａ变为Ｇ的天然存在的不表达的Ｃ３（２）等位基因上此片段的核蛋白结合能力作比较后提示在Ｃ３基因唯一表达的组织—腹侧前列腺中，结合在此区的核蛋白是－２９位Ａ依赖的，而在所研究的不表达组织—肝和睾丸中则否。而且此类蛋白质还可能具有与雄激素调控此基因表达有关的其他反式作用因子相互作用的结构域．     

小麦高分子量谷蛋白基因转录调控相关的顺反式相互作用的初步研究

利用凝胶延滞（ｇｅｌｒｅｔａｒｄａｔｉｏｎ）分析技术, 研究了小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ ）高分子量谷蛋白基因转录启始点上游９００ ｂｐ 的ＤＮＡ 片段与未成熟小麦胚乳细胞核因子的顺反式相互作用。利用ＤＮＡ聚合酶链反应技术, 将９００ ｂｐ 分成４ 段凝胶延滞分析用探针, 并以高盐浓度抽提法从扬花１２ ｄ 左右的小麦胚乳得到了核抽提物。凝胶延滞分析结果表明, 这４ 个ＤＮＡ片段上均存在与各自核蛋白因子专一性结合的位点。推测该基因的表达是受多位点复杂的顺反式相互作用来调控的     

大鼠诱导型一氧化氮合酶基因转录调控区的克隆与鉴定

采用单特异引物ＰＣＲ克隆法,得到大鼠诱导型一氧化氮合酶（ｉＮＯＳ）基因转录调控区ＤＮＡ片段．核酸序列分析证实,大鼠ｉＮＯＳ基因的５′－侧翼区含有ＩＦＮ－γ和ＴＮＦ－α应答元件及ＮＦ－κＢ结合位点的保守序列．这些保守序列的位置及排列显著区别于人和小鼠的ｉＮＯＳ基因．电泳迁移率改变分析（ＥＭＳＡ）表明,ＶＳＭＣ受ＩＬ－１和ＩＦＮ－γ刺激后,细胞核内产生某种可与ｉＮＯＳ基因５′－侧翼区特异结合的核蛋白因子．     

甜菜碱醛脱氢酶基因的cDNA克隆

甜菜碱醛脱氢酶基因的ｃＤＮＡ克隆马德钦,汤岚,吕文,骆爱玲,梁峰（中国科学院微生物研究所,北京１０００８０）（中国科学院植物研究所,北京１０００４４）ｃＤＮＡＣＬＯＮＥＯＦＴＨＥＧＥＮＥＯＦＢＥＴＡＩＮＥＡＬＤＥＨＹＤＥＤＥＨＹＤＲＯＧＥＮＡＳＥ￥￣...     

大肠杆菌cai DE的基因克隆与表达

从Ｅ．ｃｏｌｉ ＭＣ４１００菌体的染色体ＤＮＡ中利用鸟枪法克隆含编码肉碱消旋酶及相关因子的ｃａｉＤＥ基因片段,并经序列分析验证,由重组质粒ｐｌＸ３９３亚克隆得到ｐＤＳＷ２重组表达质粒,后者转入Ｅ．ｃｏｌｉ ＢＬ２１（ＤＥ３）菌株中是丙基硫代半乳糖苷（ＩＰＴＧ）诱导,在聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）上分子量为３０ｋＤ和２４ｋＤ附近可见明显的表达蛋白带。     

福尔马林固定云南鲴的DNA提取及其细胞色素b基因序列分析

福尔马林固定云南鲴的ＤＮＡ提取及其细胞色素ｂ基因序列分析ＤＮＡＥＸＴＲＡＣＴＥＤＦＲＯＭＦＯＲＭＡＬＩＮ－ＦＩＸＥＤＸｅｎｏｃｙｐｒｉｓｙｕｎｎａｎｅｎｓｉｓＡＮＤＳＥＱＵＥＮＣＥＡＮＡＬＹＳＩＳＯＦＩＴＳＣＹＴＯＣＨＲＯＭＥＢＧＥＮＥ关键词福尔马林...     

雄激素受体与雄激素应答元件的相互作用

将雄激素受体（ＡＲ）的ｃＤＮＡ１１１９ｂｐ片段（１１０５－２２２４）（其中包含其ＤＮＡ结合结构域到部分激素结合结构域）克隆于表达南粒ｐＧＥＸ中,通过ＩＰＴＧ诱地,在Ｅ．ｃｏｌｉ中表达ＧＳＴ－ＡＲ融合蛋白,经谷胱甘肽－Ｓｅｐｈａｒｏｓｅ－４Ｂ亲和层析得以部分纯化。利用一个有雄激素应答元件（ＡＲＥ）活性的Ｃ３（１）ＤＮＡ片段为阳性探针,通过凝胶阻滞分析和ＤＮａｓｅ１足迹法证明此表达产物具有牧民的ＤＮＡ     

应用放射性自显影技术检测外源DNA与鸡精子的结合

应用放射性自显影技术检测外源ＤＮＡ与鸡精子的结合ＤＥＴＥＣＴＩＮＧＡＳＳＯＣＩＡＴＩＯＮＯＦＥＸＯＧＥＮＯＵＳＤＮＡＷＩＴＨＣＨＩＣＫＥＮＳＰＥＲＭＵＳＩＮＧＡＵＴＯＲＡＤＩＯＧＲＡＰＨＹ关键词鸡，精子，脂质体，ＤＮＡ与精子的结合ＫｅｙｗｏｒｄｓＣｈ...     

不同性别表型黄瓜基因组中雌性系特异的ACC合酶基因

利用一对引物（引物１和引物２）分别从雌性系黄瓜（Ｃｕｃｕｍｉｓ ｓａｔｉｖｕｓ Ｌ．）品种“ＣＯＲＯＮＡ”、“ＤＡＬＥＶＥ”和强雌性黄瓜品种“中农五号”、“欧洲八号”的基因组ＤＮＡ中扩增到一长约１０２５ｂｐ的ＡＣＣ合酶基因片段。序列分析表明：该基因片段与Ｔｒｅｂｉｔｓｈ等１９９７年发表的ＡＣＣ合酶基因片段的同源性大于９９％,认为这两个基因片段应该是同一个基因,不同品种来源的该基因的相同性说明了其高     

Incorporation of radioactivity from labeled Di-(2-ethylhexyl)phthalate into DNA of rat liver in vivo

Di-(2-ethylhexyl)phthalate (DEHP), when fed at high levels in the diet for two years, is reportedly an hepatocarcinogen to rats and mice. Radioactivity from ethylhexyl-labeled, but not from phthalate-labeled, [14C]-DEHP is associated with highly purified DNA from the livers of treated rats and this radioactivity is not accounted for by assumptions of adsorption, intercalation, attachment to RNA or histones, an impurity in the labeled DEHP, or artifactual binding during sample workup. Spontaneous binding of radioactivity to DNA from either ethylhexyl-labeled DEHP or its total urinary metabolites could not be detected. Although rat liver slices generated all of the known metabolites of DEHP in vitro, no binding to DNA occurred. Administration of dual 3H/14C-labeled DEHP to rats yielded liver DNA whose 3H/14C ratio was inconsistent with the attachment of any reasonable multi-carbon fragment from the ethylhexyl portion to the DNA. The observation that roughly 100 times as high a percentage of the 14C administered was found in urea as in total DNA suggests that the 14C entered DNA through carbamyl phosphate, a precursor of both urea and pyrimidine bases. If this is the case, the association of C-1 from the ethylhexyl portion of DEHP with DNA may not involve alteration of the DNA or genetic damage.     

Sequence motif in control regions of the H+/K+ ATPase alpha and beta subunit genes recognized by gastric specific nuclear protein(s).

A nuclear protein(s) from rat or pig stomach recognized a conserved sequence in the 5'-upstream regions of the rat and human H+/K(+)-ATPase alpha subunit genes. A gel retardation assay suggested that part of the binding site was located in the TAATCAGCTG sequence. No nuclear proteins capable of the binding could be detected in other tissues of rat (liver, brain, kidney, spleen and lung) or pig liver. The sequence motif (GATAGC) located 5'-upstream of the beta-subunit gene also seemed to be recognized by the same protein, because the binding of nuclear protein to the sequence motifs in the alpha and beta subunits was mutually competitive. Considering the sense-strand sequence of the binding motif in the alpha-subunit gene, we conclude that (G/C)PuPu(G/C)NGAT(A/T)PuPy is a core sequence motif for the gastric specific DNA binding protein (PCSF, parietal cell specific factor).     

Binding of a nuclear factor to a consensus sequence in the 5' flanking region of zein genes from maize

The genomic organization of the zein structural genes and of regulatory loci influencing their expression suggests that control of zein gene expression will involve interactions between cis elements in the flanking DNA sequences and products from trans-acting genes. The interaction between fragments from the 5' flanking region of a zein gene and specific, double-stranded oligonucleotides with crude nuclear extracts from maize endosperm have been studied by nitrocellulose filter binding, gel retention and DNase I footprinting assays. Specific binding of a nuclear factor was observed and the exact position of the protein binding site was determined. The 22-nt binding site included 14 bp of a 15-bp sequence conserved in all zein genes.     

鸡烟碱样乙酰胆碱受体γ亚基启动子中的M-CAT结合特性(英文)

鸡骨骼肌烟碱样乙酰胆碱受体 ( ACh R)γ亚基基因启动子含有几个 M- CAT元件 ,为γ亚基基因表达其全能活性所必需 .为探索 M- CAT元件结合功能特性 ,分析了 γ亚基启动子近转录起始点 - 67/ - 61位的 M- CAT元件与核因子的相互作用 .突变及结合试验证明 ,M- CAT核心序列结合几种测试组织中的核蛋白产生高迁移的 DNA-蛋白质复合物 ;侧翼序列结合肝以外的几种组织不同的核蛋白质 ,产生低迁移的 DNA-蛋白质复合物 .Southwestern印迹 ( DNA印迹 )技术在 1 3d鸡胚所有被检测的组织核抽提物中只检出了 30 k D的核蛋白 .结果提示 ,30 k D因子在多种组织普遍表达 ,可直接以单体或同质二聚体形式结合 M- CAT元件 ;而结合侧翼序列的不同组织因子结合DNA时可能需依赖普遍表达的 30 k D因子的存在 .