Molecular cloning of growth hormone receptor (GHR) from common carp(Cyprinus carpio L.)and identification of its two forms of mRNA transcripts

Growth hormone receptor (GHR) belongs tothehematopoietic receptor superfamily[1]. The action ofgrowth hormone (GH) in regulating growth[2],re-production[3]and i mmunity[4]has been elucidated.The binding of GHtothe GHRontarget tissues trig-gers a cascade of tyrosine and protein phosphorylationevents, which cul minates in the biological action ofGH[5 ,6]. Up to date GHR cDNAs have been clonedfrom many species[7—9],including various kinds ofmammalian ani mals ; avian of chicken and domest…     

Transgene for growth hormone in common carp（Cyprinus carpio L.） promotes thymus development

The transgenic carp were produced by micro-injection of CAgcGHc into the fertilized eggs. Observation of the thymus development between the transgenics and non- transgenic controls was carried out. The thymus of one-year- old transgenics F1 showed a great increase in both size and weight. The unilateral thymus of the transgenics weighed from 190 to 295 mg with average 218.6 mg, whereas the uni-lateral thymus of the controls weighed 20—81 mg with av-erage 42.5 mg; i.e. the thymus weight in the transgenics was 5.14 fold over that in the controls. The index of thymus/body weight in the transgenics was 2.97 fold over the controls. Light microscopy observation indicated that the thymus of the transgenics well developed with the thickened outer re-gion and compactly arranged thymocytes, while the thymus in the controls were degenerating with the thinned outer region, scattered thymocytes and groups of fatty cells. Fur-ther analysis with the electron microscopy revealed that pro-liferous cells in the transgenics were mainly small lympho-cytes and no pathological changes were found. The results confirmed that the 揂ll-fish?GH-transgene promotes thy-mus development and thymocyte proliferation, and retards thymus degeneration. The study has laid a foundation for further analysis of the immunobiological function in GH- transgenic carp.     

胞内表达猪生长激素基因的重组酵母菌构建与鉴定

将猪生长激素(pGH)cDNA定向插入毕赤酵母胞内表达载体pPIC 3.5k质粒,重组构建了pPIC 3.5kpGH/GS115酵母表达系统, 其表达产物的SDSPAGE和Western blot结果表明, pGH蛋白在胞内获得正确表达,表达量达378 mg/L.     

人表皮生长因子(hEGF)基因的合成、鉴定及植物表达质粒的构建

用PCR方法合成了人表皮生长因子（hEGF）基因，构建了原核表达质粒p 20T-hEGF，研究了原核表达的重组蛋白hEGF的生物学活性，并构建了植物表达质粒，研究表明：无论是融合蛋白GST－hEGF或纯化的hEGF蛋白，都有相当高的生物活性，hEGF蛋白对Hela细胞的增殖有很好的促进作用，免疫小鼠亦能产生很高的免疫应答反应。     

草鱼肌肉生长抑制素cDNA克隆及序列分析

利用RT-PCR技术,从草鱼肌肉组织中克隆出MSTN cDNA序列,长度为1764 bp,编码区为1128 bp,共编码375个氨基酸.前体蛋白包括信号肽序列、N端前肽区、蛋白酶水解位点RIRR及C端活性区(含9个保守的Cys残基).多重序列比较表明,草鱼与斑马鱼、黑头软口鲦之间的MSTN序列同源性在94%以上,与其它鱼类之间的序列同源性也较高(78.1%~82.3%),但与鸟类、哺乳类之间的序列同源性则相对较低.系统进化分析显示,不同物种间的MSTN序列同源性基本反映了它们的亲缘关系.     

鲤NADH泛醌氧化还原酶亚基3基因的克隆及低温适应相关性分析

根据鲤低温下特异表达的基因（片段）序列设计引物,采用2轮菌落PCR方法对鲤脑全长cDNA文库中的克隆进行筛选,确定了基因序列152在文库中的位置,经生物信息学比对分析确定其为NADH泛醌氧化还原酶亚基3基因.这一基因在低温下的特异表达,对于低温下鱼类机体的供能,起着重要作用.结果还表明,NADH泛醌氧化还原酶亚基3基因在鲤氧化呼吸链电子传递系统中与低温适应性状在一定程度上存在相关性.     

大肠杆菌中重组猪生长激素提取方法及变复性的比较研究

通过对基因工程菌pgh／E．coli BL21用酶溶解、超声波处理、超声波处理与液氮冻融相结合三种方法进行破菌，用盐酸胍法进行变复性，提取重组猪生长激素(rpGH)．结果表明用超声波处理与液氮冻融相结合的方法裂解细菌，盐酸胍法变复性，rpGH的提取率最高，达72．16％；而用尿素法对rpGH在大肠杆菌内所形成的包涵体变复性，rpGH的提取率仅为49．82％．     

Construction of a BAC library from cucumber (Cucumis sativus L.) and identification of linkage group specific clones

A bacterial artificial chromosome (BAC) library consisting of 19,200 clones with an average insert size of 105 kb has been constructed from a cucumber (Cucumis sativus L.) inbred line S94; derived from a cultivar in North China. The entire library was equivalent to approximately 5 haploid cucumber genomes. To facilitate chromosome engineering and anchor the cucumber genetic linkage map to its chromosomes, 15 sequence-characterized amplified regions (SCAR) and seven simple sequence repeats (SSR) markers from each linkage group of cucumber were used to screen an ordered array of pooled BAC DNA with polymerase chain reaction (PCR). Fifteen markers gave at least two positive clones. As a result, 32 BAC clones representing 7 linkage groups of cucumber were identified, which further validated the genome coverage and utility of the library. This BAC library and linkage group specific clones provide essential resources for future research of the cucumber genome.     

Construction of a BAC library from cucumber (Cucumis sativus L.) and identification of linkage group specific clones

A bacterial artificial chromosome (BAC) library consisting of 19,200 clones with an average insert size of 105 kb has been constructed from a cucumber (Cucumis sativus L.) inbred line S94; derived from a cultivar in North China. The entire library was equivalent to approximately 5 haploid cucumber genomes. To facilitate chromosome engineering and anchor the cucumber genetic linkage map to its chromosomes, 15 sequence-characterized amplified regions (SCAR) and seven simple sequence repeats (SSR) markers from each linkage group of cucumber were used to screen an ordered array of pooled BAC DNA with polymerase chain reaction (PCR). Fifteen markers gave at least two positive clones. As a result, 32 BAC clones representing 7 linkage groups of cucumber were identified, which further validated the genome coverage and utility of the library. This BAC library and linkage group specific clones provide essential resources for future research of the cucumber genome.     

水稻蔗糖转化酶基因的克隆及其功能的初步探讨

根据抑制消减杂交(suppression subtractive hybridization，SSH)方法分离到在水稻叶片中高表达的蔗糖转化酶基因片段，根据水稻基因组顺序设计引物分离到全长cDNA．测序结果表明该cDNA长度为1937bp，推测编码一个627个氨基酸的中性/碱性蔗糖转化酶．Alignment分析显示该推测蛋白质与已知的多个蔗糖转化酶具有很高的同源性．半定量PCR检测证实它在叶中的表达强于在根、花药和幼穗等组织中的表达，胁迫处理(盐和低温)使其表达上调．     

菜芙蓉总黄酮纯化及其体内抗氧化性

为研究菜芙蓉总黄酮(TFA)的纯化条件及体内抗氧化性,采用AB-8大孔树脂静态吸附-洗脱工艺,控制纯化条件为上样液质量浓度0.1g/mL,pH值为3.5,解吸液乙醇体积分数为70%,解吸液体积与树脂质量比为20∶1(mL/g)。在最优纯化条件下,黄酮的质量分数提高到48.50%;与正常对照组相比,实验组小鼠器官指数无显著差异(P0.05);小鼠血清和肝组织中MDA含量显著低于正常对照组(P0.05),而CAT,GSH-Px和SOD活性均高于正常对照组且具有明显的量效关系。实验结果表明,TFA具有良好的体内抗氧化效果。     

Conditional and unconditional mapping of quantitative trait loci underlying plant height and tiller number in rice (Oryza sativa L.)grown at two nitrogen levels

In this study, we used 127 double haploid (DH) lines to analyze agricultural traits of rice. The DH lines, derived from a ZYQ8 (indica)/JX17 (japonica) cross by anther culture, contained 160 RFLP and 83 SSR markers. Unconditional and conditional quantitative trait loci (QTL) mapping was conducted to analyze plant height (PH) and tillers per plant (TP) at ?ve growth stages that were grown at two nitrogen levels. Fourteen PH and 13 TP unconditional QTL were identified in the di?erent growth stages, including 19 QTL from high-nitrogen (HN) and 14 QTL from low-nitrogen (LN) conditions. The conditional QTL for 14 genomic regions under LN/HN conditions showed that there was a significant effect on PH and TP across the different stages. Only one conditional QTL, ph2-3, was unable to be detected in unconditional mapping. More QTL were detected in the ?rst four rice growth stages than in the final stage. Furthermore, a line from the DH mapping population, DH78, was identified in extreme phenotypes of PH and TP that exhibited dwarfism and less-tiller (dft) characters. The gene dft1 was mapped to chromosome 2 using a backcrossed population of DH78/JX17 through a mapbased cloning strategy. The location of dft1 coincided with the mapping region of the small-LOD peak, QTL ph2 and tp2, which were identified in plants grown in low-nitrogen conditions. Further backcrossing and fine-mapping successfully delimited the dft1 locus to a 91 kb region.     

鸡粪嫌气发酵液毒害植物的有机成分及其对莴苣种子萌发生理影响

气相色谱分析的结果表明,由活性碳分离出来的鸡粪嫌气发酵液（ADCME）对植物产生毒害的主要有机化合物是异戊酸,丁酸和异丁酸,分别占45％,35％和20％,用生物分析来鉴定ADCME的活性碳提取物和这三种挥发性脂肪酸及其混合物对莴苣（Lactuca sativa L.),种子萌发,根芽生长和幼苗中的生化活动的效应。生化分析表明这些脂肪酸和ADCME提取物均抑制莴苣贮藏蛋白质的重新利用,导致其对Mg^2 -ATP酶,Ca2 -ATP酶,多酚氧化酶和过氧化物酶活性以及ATP酶的作用产物之一的无机Pi的释放都受到显著的抑制,这三种脂肪酸中,戊酸处理的抑制作用最强,丁酸处理次之,并观察到有机混合物处理具有协和效应。