赛默飞展示端到端的生物样本库和生物储存库解决方案

整体式生物样本库解决方案保障了生物样本的质量,提高了效率并促进了信息共享 中国上海,2011年11月16日——全球科学服务领域的领导者赛默飞世尔科技（以下简称：赛默飞）于11月16-19日期间在法国马赛举行的ESBB2011上展示其整体式生物样本库解决方案。这一综合性解决方案保障了生物样本质量,并涵盖了完整的生物样本库工作流程,包括样本收集、处理、     

A new combination of mutated loxPs in a vector for construction of phage antibody libraries

In the construction of large antibody libraries by in vivo recombination, two non-homogeneous loxP sites are required for the exchange of Vgenes between phagemids to create many new VH-VL combinations.The mutated loxP511 was designed not to recombine with the wild-type loxP (loxPwt) in early studies and a combination of the two has been used to construct antibody libraries. But recent reports have shown that recombination occurs between loxPwt and loxP511. This suggests that the combinational use of loxP511 and loxPwt might lead to the loss of the V gene diversity of antibody libraries. Therefore, it is necessary to find a new combination of loxPs to avoid the excision recombination in the antibody library. In this study,we found that the excision recombination between loxP511 and loxP2272, another mutated loxP sequence,was undetectable within one phagemid, while the excision recombination between loxP511 and loxPwt occurred at a frequency of 40%, higher than that reported previously. Furthermore, the in vivo recombination of different phagemids with loxP511 and loxP2272 showed that the V gene exchange was efficiently mediated to produce new VH-VL combinations. It was concluded that the loxP511 and loxP2272 combination was more favorable for reducing the excision recombination and constructing large phage antibody libraries with high diversity.     

抗体工程研究进展

噬菌体抗体库技术的出现为抗体工程的研究带来了革命性的变化,本文总结了当前国内外抗体工程,特别是单链抗体方面的研究进展。     

从噬菌体随机肽库中筛选流感病毒神经氨酸酶的抑制多肽

目的:从噬菌体呈现12肽库中筛选与流感病毒神经氨酸酶特异性结合的肽。方法:以甲三型流感病毒裂解疫苗原液为靶分子,经过3轮生物淘选,从噬菌体随机肽库中筛选与之结合的噬菌体。用ELISA方法鉴定噬菌体克隆与靶分子的结合力,用荧光方法测定噬菌体克隆对流感病毒A/Sydney/5/97(H3N2)神经氨酸酶的抑制活性。对筛选到的阳性克隆进行DNA序列测定并推导出相应的氨基酸序列。结果:经过3轮筛选后,42个噬菌体克隆与靶分子有高度亲和力,23个噬菌体克隆对流感病毒A/Sydney/5/97(H3N2)神经氨酸酶有抑制活性。对27个噬菌体克隆的测序结果表明,分别有10个和2个克隆的序列是一致的,其氨基酸序列分别为KSLSRHDHIHHH和WPRHHHSASVQT。结论:通过噬菌体肽库筛选到抑制流感病毒神经氨酸酶的12肽,为进一步研究对流感病毒神经氨酸酶有抑制活性的分子药物奠定了基础。     

Construction and Characterization of a cDNA Library from the Pulp of Cara Cara Navel Orange （Citrus sinensis Osbeck）

A cDNA library was constructed end characterized from the pulp of Cera Care navel orange （Citrus sinensis Osbeck） at different stages of ripening. Tittering results revealed that approximately 5.086×10^5 independent clones were included in this library. Electrophoresls gel results of 15 randomly selected clones revealed that the size of the insertion fragments ranged from 400 bp to 2 kb, with an average size of 900 bp. Sequencing results of 150 randomly picked clones showed that the recombination rate was 94%. During subsequent sequence analysis, 41 of 139 clones failed to be identified end the amino sequence of 71 clones shared less than 30% identity with related plants in GenBank. Of 27 clones whose amino sequences shared more than 60% identity with other related plants in GenBenk, 17 clones showed an 80% identity with the corresponding candidate genes of citrus. The clone recognized as the type Ⅲ metallothlonein-llke （MT） gene was observed to occur 13 tlmes, Indlcatlng that the protein may play an important role In frult development and rlpenlng.     

基于源-库互反馈的温室青椒坐果时空动态模拟

基于源-库互反馈的温室青椒坐果时空动态模拟     

种植密度对夏玉米产量和源库特性的影响

以高产玉米品种郑单958(ZD958)和登海661(DH661)为试验材料,在4个不同区域(山东农业大学、汶口、兖州和莱州)设置22500、45000、67500、90000和112500株.hm-25个种植密度,研究了种植密度对夏玉米产量及源库特性的影响.结果表明:两品种在112500株.hm-2密度条件下玉米籽粒产量和生物产量最高,分别为19132和36965kg.hm-2,与22500和67500株.hm-2密度相比,籽粒产量分别增加了72%和48%,生物产量分别增加了152%和112%.两品种单株叶面积、最大花丝数、穗粒数和千粒重随密度增大而减小,但叶面积指数随密度增大而显著提高.收获指数与粒叶比随密度增大而显著减小,当密度超过67500株.hm-2时差异不显著,表明高密度条件下玉米通过增加群体库来提高产量.     

农杆菌介导的紫色红曲霉遗传转化体系的建立和优化

通过优化各种转化因素,建立了根癌农杆菌(Agrobacterium tumefaciens)介导红曲霉(Monascus)的高效转化体系:红曲霉在PDA培养基培养21 d后收集孢子,制备红曲霉孢子悬浮液,浓度为106个/mL,根癌农杆菌浓度为OD600值0.5,诱导剂AS浓度为100μmol/L,农杆菌与红曲霉在25℃共培养3 d。采用此转化体系构建了含有530多个转化子的红曲霉T-DNA插入突变体库。随机选取50株转化子菌株进行分子验证和稳定性检测,证明T-DNA成功插入红曲霉基因组DNA中,并能稳定遗传。最后,通过形态观察筛选出8株变异较大的菌株,为以后的红曲霉基因功能研究奠定了一定的基础。     

顺乌头酸酶基因在杀雄剂SQ-1诱导小麦雄性不育花药中的表达分析

采用RT-PCR技术,克隆了小麦胞质顺乌头酸酶基因(cACO)部分cDNA序列.该cDNA序列长1368bp,编码456个氨基酸,GenBank登录号为GU475062.半定量RT-PCR结果表明,在生理型不育和可育花药发育的单核早期至三核期,cACO基因的表达水平均表现为先升后降;在生理型不育花药发育的单核晚期cACO基因表达水平与同期可育花药相比显著升高,到二核期和三核期明显降低,ACO酶活性变化表现出相同趋势.这反映出在小麦生理型不育系中,cACO基因在花药败育关键期异常表达可能影响了花药发育过程中正常的能量供应和物质代谢,导致花粉发育能量不足和所需物质匮乏,从而导致了非遗传型花药败育现象.     

噬菌体抗体库技术及其应用研究进展

噬菌体呈现抗体库是近年发展的一项分子生物学新技术,它的建立是抗体技术领域中的一次革命性进展。它以其独特的构建和筛选系统,彻底改变了抗体制备的传统途径,使抗体工程技术进入了一个新的发展阶段,并对生物学领域中许多技术的发展起到了巨大的推动作用。该技术是迄今发展最成熟、应用最广泛的制备抗体技术。我们简要综述此项技术的研究应用进展。