重组FAS分子的表达、纯化及抗体制备

用ＰＣＲ法扩增出编码人ＦＡＳ分子胞外区的ｃＤＮＡ片段,直接克隆到ｐＧＥＭ－Ｔ载体上,经ＤＮＡ序列测定后,再插入到谷胱甘肽转硫酶（ＧＳＴ）融合蛋白表达载体ｐＧＥＸ－ＫＧ的ＥｃｏＲⅠ和ＳａｌⅠ位点之间,构成重组质粒ｐＫＧ－ｈＦＡＳ,将此质粒导入大肠杆菌,经ＩＰＴＧ诱导后获得ＧＳＴ－ｈＦＡＳ重组融合蛋白的表达,用谷胱甘肽偶联的Ｓｅｐｈａｒｏｓｅ４Ｂ经亲合层析获得纯化的ＧＳＴ－ｈＦＡＳ蛋白,经凝血酶酶切和二次亲合层析去除ＧＳＴ部分,得到纯化的ＦＡＳ蛋白．用纯化的ＦＡＳ抗原免疫家兔制备了抗ＦＡＳ抗体,经检测发现抗ＦＡＳ抗体能诱导Ｕ９３７细胞发生细胞凋亡     

转化生长因子α—绿脓杆菌外毒素融合蛋白TGFa—PE40高效表达质…

利用基因工程技术,将质粒ＰＹＸ３８２用ＸｂａＩ和ＥｃｏＲＩ切下插入的ＴＧＦａ－ＰＥ４０融合基因片段,连接到可表达载体ＰＣＢ６０４的ＸｂａＩ／ＥｃｏＲＩ位点中,构建成新的重组质粒Ｐ２Ｘ－ＴＰ１。Ｐ２Ｘ－ＴＰ１转化Ｅ．ｃｏｌｉＢＬ２１感受态菌后,在ＩＰＴＧ诱导下Ｉｐｐ启动子转寻表达ＴＧＦａ－ＰＥ４０融合蛋白。表达产生物主要以包涵体形式沉积在细胞内,隔合蛋白表达量的高低与诱导时的细胞密度,诱导的温度以及     

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     

外源TGF—β1基因在小鼠ES细胞中的表达及其对细胞分化的影响

本文利用逆转录病毒载体Ｄｏｌ,在其ＢａｍＨＩ酶切位点插入猪ＴＧＦ－β１ １．７ｋｂｃＫＮＡ,构建成表达质粒,并用磷酸钙沉淀法将该质粒ＤＮＡ转染到小鼠ＥＳ－５细胞,经Ｇ４１８筛选获得抗Ｇ４１８的ＥＳ－５细胞克隆（ＥＳ－Ｔ）,经ＲＮＡ点杂交,Ｎｏｒｔｈｅｒｎ印迹杂交证明有６个细胞克隆能表达外源猪ＴＧＦ－β１的ｍＲＮＡ,其中两个杂交信号较强的克隆进一步用Ｓｏｕｔｈｅｒｎ印迹杂交,也证明猪ＴＧＦ－μ     

水稻齿叶矮缩病毒Segment 9 dsRNA的序列分析及其在大肠杆菌DE3中的表达

通过有机试剂抽提,ＣＦ－１１纤维素柱层析,从感染水稻齿叶矮缩病毒菲律宾分离株（ＲｉｃｅＲａｇｇｅｄＳｔｕｎｔＶｉｒｕｓ,Ｐｈｉｌｉｐｐｉｎｅｉｓｏｌａｔｅ,简称ＲＲＳＶ－Ｐ）的水稻植株中获取该病毒的全基因组,即获得从Ｓｅｇｍｅｎｔ１到Ｓｅｇｍｅｎｔ１０（Ｓ１－Ｓ１０）的１０条双链ＲＮＡ（ｄｓＲＮＡ）,然后设计合适的引物,用ＲＴ－ＰＣＲ方法得到Ｓ９的ｃＤＮＡ并将其克隆到ｐＵＣ１１９质粒上扩增,以双链测序法测定该ｃＤＮＡ的全序列。同时又将此ｃＤＮＡ克隆到大肠杆菌表达质粒ｐＧＥＸ－３Ｘ上,在大肠杆菌菌株ＤＥ３中用ＩＰＴＧ诱导表达,经超声波破菌、离心、Ｇｌｕｔａｔｈｉｏｎｅ－ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析,得到纯化的分子量为６４ｋＤ的融合蛋白。     

人表皮生长因子(EGF)与单核-巨噬细胞集落刺激因子(GM-CSF)融合蛋白在大肠杆菌中的表达

应用基因工程技术,将ＥＧＦ、ＧＭ－ＣＳＦ基因克隆到ｐＧＥＭ－３Ｚｆ（＋）载体的ＥｃｏＲＩ,ＢａｍＨＩ位点上,再将重组融合基因亚克隆到表达载体ｐＢＶ２２０的ＥｃｏＲＩ,ＢａｍＨＩ位点上,在大肠杆菌ＤＨ５α中进行表达,ＳＤＳ－聚丙烯酰胺凝胶电泳和Ｗｅｓｔｅｒｎｂｌｏｔ表明ＥＧＦ－ＧＭ－ＣＳＦ融合蛋白获得表达,并且具有ＥＧＦ、ＧＭ－ＣＳＦ的免疫学活性．这为进一步研究该融合蛋白的功能和肿瘤治疗提供一种新的基因产品．     

人睫状神经营养因子基因及突变体在大肠杆菌中的表达

人睫状神经营养因子（ｈｕｍａｎ ｃｉｌｉａｒｙ ｎｅｕｒｏｔｒｏｐｈｉｃ ｆａｃｔｏｒ,ｈＣＮＴＦ）ｃＤＮＡ克隆入具有ＰＬ启动子的表达载体,转化大肠杆菌ＨＢ１０１,构建成表达ｈＣＮＴＦ菌株。热诱导后,ｈＣＮＴＦ的表达量达菌体总蛋白量的２５％。用离子交换层析、凝胶过滤层析从菌体裂解液中纯化了ｈＣＮＴＦ。ＳＤＳ－ＰＡＧＥｈＣＮＴＦ的分子量约为２４ｋｄ,Ｎ端氨基酸序列分析结果与依基因核苷酸推导疗列相符。     

转录因子NF-κB p50基因的克隆、表达及抗p50抗体的制备

利用ＤＮＡ重组技术,将转录因子ＮＦ－κＢｐ５０基因片段插入ｐＧＥＸ－２Ｔ质粒载体,经ＤＮＡ序列分析证明克隆的正确性．将重组的ｐＧＥＸ－２Ｔ／ｐ５０转化入大肠杆菌ＪＭ１０９,表达出ｐＧＥＸ－２Ｔ／ｐ５０融合蛋白．经ＷｅｓｔｅｒｎＢｌｏｔ检测证实表达产物对抗ｐ５０ｃ的抗体呈特异性反应,表明克隆的正确性,并成功地应用纯化的ｐ５０蛋白制备了免疫血清．为进一步对ＮＦ－κＢ的功能进行研究打下了良好的基础     

BIV—1—gp120在重组杆状病毒系统中的表达

将中国株ＨＩＶ－１Ｂ亚型ｇｐ１２０全基因序列克隆到杆状病毒转座载体ｐＦａｓｔＢａｃＩ中多角体启动子下游,构建成重组转座载体ｐＦａｓｔＢａｃＩ－ｇｐ１２０,利用细菌／杆状病毒（Ｂａｃ ｔｏ Ｂａｃ）表达系统筛选重组杆状病毒,在昆虫细胞Ｓｆ９中高效表达了ＨＩＶ－１的外膜糖蛋白ｇｐ１２０,ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎ ｂｌｏｔ分析结果一致,证明表达了２种糖基化程度不同的ｇｐ１２０。     

人表皮生长因子（EGF）与单核—巨噬细胞集落刺激因子（GM—CSF）融合？…

应用基因工程技术,将ＥＧＦ,ＧＭ－ＣＳＦ基因克隆到ｐＧＥＭ－３Ｚｆ载体的ＥｃｏＲＩ,ＢａｍＨＩ位点上,再钭重组融合基因亚克隆到表达载体ｐＢＶ２２０扔ＥｃｏＲＩ,ＢａｍＨＩ位点上,在大肠杆菌ＤＨ５α中进行表达,ＳＤＳ－聚丙烯酰胺凝胶电泳和Ｗｅｓｔｅｒｎ ｂｌｏｔ表明ＥＧ－ＦＧＭ－ＣＳＦ融全蛋白获得表达,并且具有ＥＧＦ,ＧＭ－ＣＳＦ免疫学活活。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.