We do not know what cell cycle - associated proteins are regulated by histone acetylation modification , especially at the checkpoints of g1 / s , s / g2 , g2 / m and mitosis exit . neither do we know about the manner in which histone acetylation regulates the expression of these genes 在真核细胞周期调控的重要检验点g1 / s和g2 / m ，以及其他的转换点s / g2和走出m期的各个过程中，哪些细胞周期调控相关蛋白的表达活性与细胞内的组蛋白乙酰化水平相关
Histon acetylation plays an important role in regulation chromatin structure and transcriptional activity . however , there is a lit tle investigation in cloned cattle about histone acetylation . andmore , in many dead cloned cattle , the lung abnormality is mostly obvious 在报道的克隆牛发育异常中，肺脏的异常比较普遍和明显，而在我们的克隆牛工作中也发现了类似问题，所以我们选用克隆牛肺脏组织作为实验材料，进行组蛋白乙酰化研究。
2 , by using the above system , we demonstrated that yelp3 , yhelp3 and help3 were able to completely , dramatically and partially complement the growth defects and the slow activation of pho5 and ssa3 gene caused by the depletion of yelp3 , respectively , while yhelpbhat " whose catalytic domain was partially de 7 、 h4ks一r对i功五艺p3 、力几p3与少凡p3功能的影响大于h3k14一r ，表明体内组蛋白不同位点的乙酞化修饰对基因表达有不同的影响。
In order to investigate the histone acetylation in gene different regions , we used normal cattle 4n1 , 6n1 , 9n3 , cloned cattle xiaobai and 9c4 , with the igf1 and gh genes . we designed 14 and 8 pairs of primers on their dna sequences , respectively 为研究基因不同区域的组蛋白乙酰化水平，我们选取了正常牛4n1 ， 6n1 ， 9n3 ，克隆牛xiaobai和9c4共五个样品， igf1和gh两个基冈作为研究对象，在它们的基因序列上分别设计了14对和8对引物来进行定量分析。
After washing with reagent , adopt the newest purification technology source30rpc , sds - page and densitometric scan analysis , the result show that expression level is 90 % of total bacterial proteins . after renaturation , ifnr , hgfa , hgfb , hpk5 were purified by akta purifier chromatogram instrument , sepharose fast flow , ssphacrayl series gel , selecting optimize condition . finally establish a kind of high efficient purification model of recombinant proteins produced in escherichia coli as inclusion bodies , purification product purity > 98 % 结论:总之，通过对发酵罐中重组工程菌各种培养因素的研究，建立了一种高密度、高表达发酵工艺体系，为重组蛋白的后续纯化提供了大量、稳定的原料供应;通过对不同目的蛋白的色谱行为的系统研究，建立了一种高效稳定、快速简洁、易于放大的包涵体重组蛋白分离纯化体系。
2 , the hsp gene transcription was quantitatively determined by rt - pcr . based on this result , it is concluded that the change of acetylation level at the loci of hsp , mediated by histone deacetylase inhibitors , exerts important functions in hsp gene transcription . 3 , after immunolabeling with anti acelated - lysine monoclonal antibody on the polytene chromosomes of heat shocked flies , fluorescence signals were detected at the hsp loci 2 ，利用rt - pcr技术，对去乙酰化酶抑制剂处理后的果蝇的热休克基因的表达水平进行检测，结果表明经组蛋白去乙酰化酶抑制剂处理后的果蝇幼虫，其热休克基因的表达高于基础水平，也就是说去乙酰化酶抑制剂诱导了热休克基因的表达。
This subject uses the genetic engineering technology and the newest modern biotechnology to study the genetic engineering lower reaches stage , in order to research and establish the optimum , high density fermentation technology pattern and high efficient isolation and purification model of recombinant proteins from inclusion bodies 本课题利用基因工程技术，重点从基因工程下游阶段着手研究，利用最新现代生物技术研究并建立经优化的高密度发酵工艺模式及高效分离纯化包涵体重组蛋白模式。一
Epigenetic modification of the genome ensures proper gene activation during development and involves : genome methylation changes ; the assembly of histon and histone variants into nucleosomes and remodeling of other chromatin - associated proteins such as linker histones , polycomb group , nuclear scaffold protein , and transcription factors 这些染色质的再程序化包括dna的甲基化和核小体组蛋白的乙酰化等。组蛋白乙酰化在染色质结构调节、 dna复制、基因转录、细胞分化及癌细胞发育都有研究，但在克隆动物中却是一个有待深入探索的领域。
Notch interaction with its ligands induces the cleavage of its intracellular domain ( ic ) , and the notch ic translocates to the nucleus and binds to rbp - j , the mammalian homolog of su ( h ) , to transactivate transcription of target genes such as e ( spl ) ( enhancer of split ) , hesl ( hairy and enhancer of split ) and hes5 four notch receptors and their ligands are differentially and redundantly expressed in a variety of vertebrate tissues 它通过其识别序列（ cogtgggaa ）结合于受调控基因的启动子区，在转录激活因子的驱动下调节细胞分化和个体发育相关基因的表达。在没有n 。 tch胞内段的情况下， rbpj可与包含sm盯（ silencingmediatorforretlnoldandthyroidhormonereceptor ）和组蛋白去乙酞化酶的转录辅助抑制复合物结合，当notch信号被激活时； rbpj可与n 。
In this study , the recombinant plasmid pmd - 18t - pea - h3 was cleavaged with ncoi , xhoi and inserted into the expression vector pet - 28c and subsequently subjected to restriction endonuclease analysis and sequencing , the result indicated that the prokaryotic expression vector pet - 28c - pea - h3 was constructed successfully . after the expression plasmid was extracted and transformed into expression hosts bl21 ( de3 ) of e . coli , the transformed hosts were induced by iptg , bysds - page and elisa analysis of host protein . the expression of the objective gene was detected , and it could account for 16 . 28 % of the total host protein . inclusion body was prepared from the incubating expression hosts induced by iptg 同时将原核表达载体pet - 28c用nco ， xho双酶切，回收酶切产物，将回收的酶切产物pea ， h3 ，载体进行连接，并转入dh5感受态细胞内，培养12 - 18小时后，挑取阳性菌落，经nco ， xho双酶切分析及pcr检测，筛选到阳性克隆，其质粒测序结果表明成功地构建了毒性基因缺失的pea与人组蛋白h3融合基因的原核表达载体。
