乙型肝炎病毒蛋白和绿色荧光蛋白的共表达研究

目的：构建增强型绿色荧光蛋白（EGFP）标记的乙型肝炎病毒（HBV）真核表达载体,并研究其在真核细胞和小鼠体内的共表达。方法：以质粒pBR322-HBVadr2．0和pCX-EGFP为基础,构建含有双拷贝HBV全基因组DNA和EGFP基因的真核表达载体pCX-EGFP-HBVadr2．0,分别转染真核细胞和小鼠肝组织,建立体外、体内表达系统,研究GFP和HBV基因的表达。结果：构建了真核表达载体pCX-EGFP-HBVadr2．0,EGFP和HBV病毒蛋白在体内和体外均可表达。结论：构建的pCX-EGFP-HBVadr2．0真核表达载体可以GFP作为HBV存在与否的报告基因,提高了培育检测转基因小鼠的效率,为转基因小鼠的制备及后续研究奠定了基础。     

Interspecies recombination, and phylogenetic distortions, within the glutamine synthetase and shikimate dehydrogenase genes of Neisseria meningitidis and commensal Neisseria species

Visual inspection showed clear evidence of a history of intraspecies recombinational exchanges within the neighbouring meningococcal shikimate dehydrogenase ( aroE  ) and glutamine synthetase ( glnA) genes, which was supported by the non-congruence of the trees constructed from the sequences of these genes from different meningococcal strains, and by statistical tests for mosaic structure. Many examples were also found of highly localized interspecies recombinational exchanges between the meningococcal aroE and glnA genes and those of commensal Neisseria species. These exchanges appear to have inflated the sequence variation at these loci, and have resulted in major distortions of the phylogenetic trees constructed from the sequences of the aroE and glnA genes of human pathogenic and commensal Neisseria species. Statistical tests for sequence mosaicism, and for anomalies within the Neisseria species trees, strongly supported the view that frequent interspecies recombination has occurred within aroE and glnA . The high levels of sequence variation, and intra- and interspecies recombination, within aroE and glnA did not appear to be due to a 'hitch-hiking' effect caused by positive selection for variation at a neighbouring gene. Our results suggest that interspecies recombinational exchanges with commensal Neisseria occur frequently in some meningococcal 'housekeeping' genes as they can be observed readily even when there appears to be no obvious selection for the recombinant phenotypes.     

南京市2011年乙型流感血凝素基因分子特征分析

[目的]分析2011年南京市乙型流感病毒的血凝素(HA)分子学特征.[方法]选择7株2011年南京市不同时间段有代表性的乙型流感毒株进行HA基因序列测定,通过生物信息学方法对HA分子学特征进行分析.[结果]7株乙型流感毒株分为两个系,4株为Victoria,3株为Yamagata;与2011年度疫苗株相比,Victoria和Yamagata系毒株分别在抗原位点146、197和116、198发生了氨基酸替换;其中197和198位点分别是Victoria和Yamagata毒株的受体结合位点,由于上述位点的替换使得Victoria系/Yamagata系毒株分别在197/196位增加了一个潜在的糖基化位点.[结论]2011年南京市乙型流感Victoria 系和Yamagata系病毒同时存在,Victoria/Yamagata毒株197/198位点的氨基酸替换,值得做进一步的探讨.     

西藏地区拟康蝗属一新种(直翅目：网翅蝗科）

记述采自中国西藏网翅蝗科拟康蝗属Kangacrisoides Wang et al.,2006 1新种：江达拟康蝗Kangacrisoides jiangdaensis sp. nov.。新种同霍城拟康蝗K. huochengensis Wang, Zheng & Niu, 2006近似,其区别特征为：头背面具中隆线;雄性头侧窝长为宽的2.5–3倍,而雌性为3倍;雌性前翅在背面狭分开,而雄性刚刚相连;雄性前翅顶到达第5–6腹节背板,而雌性到达第4腹节背板;雄性中脉域为肘脉域宽2倍,而雌性为3倍;后足胫节内侧之上、下距近等长;鼓膜孔宽卵形;雌性下生殖板后缘角形突出。     

多种通气策略下的高浓度乙醇生产

氧气在环境胁迫强的高浓度乙醇发酵中具有重要作用.考察了自絮凝酿酒酵母在多种通气策略下的乙醇发酵及絮凝状况.使用氧化还原电位(ORP)检测发酵液中氧浓度并划分了厌氧、微氧和好氧状态.厌氧条件下的终点乙醇浓度最低(119±1.5 g/L);微氧条件下使用ORP精密控制氧气供给取得较高的乙醇浓度(131±1.8和125±1.7 g/L);在通气量0.2 vvm的好氧条件下,生物量、甘油量和乙醇损失皆最大,与最优收率相比较乙醇收率降低了12.2％.高通气量增强了细胞的絮凝能力,增大了絮凝体粒径.绘制雷达图进行综合评价,恒定通气0.05 vvm的过程在乙醇生产和絮凝各方面表现均突出.     

牦牛BLG基因5'调控成分的克隆及序列分析

目的:克隆牦牛β-乳球蛋白(BLG)基因5'调控成分(3.5kb).方法:利用PCR方法克隆了牦牛BLG基因5'侧翼区,并进行生物信息学分析表明.结论:获得了3.5kb的BLG基因5'调控成分,其中包括5'侧翼区(2 472bp),第一外显子及第一内含子(1 066bp).该序列与牛、绵羊和山羊的同源性分别为98.42%、89.61%和84.41%;平均GC含量为49.3%;存在一个CpG岛;可能存在一个启动子位点,位于起始密码子前-83bp处;AliBaba2.1分析发现该区域有47种潜在的转录因子结合位点,其中包括乳蛋白结合因子(MPBF)、乳腺活化因子(MAF)、糖皮质激素受体(GR)、雌激素受体(ER)、核因子1(NF-1)等结合位点,提示该调控成分可用于构建乳腺特异性表达载体.结果:成功克隆出BLG基因5'侧翼区,为构建转基因牦牛乳腺生物反应器的特异性表达载体奠定基础.     

Efficient Evergreen Plant Regeneration of <i>Cinnamomum japonicum</i> Sieb. through <i>in vitro</i> Organogenesis

Cinnamomum japonicum Sieb. is an excellent roadside tree and medicinal tree species with considerable ornamental and economic value. In this study, we successfully developed a large-scale micropropagation protocol for C. japonicum for the first time. Sterilized shoots were excised and used as explants for shoot induction on several basal media, supplemented with different concentrations of plant growth regulators (PGRs), such as Thidiazuron (TDZ), N⁶ -Benzyladenine (6-benzylaminopurine) (BA), α-naphthaleneacetic acid (NAA) and Gibberellic acid (GA₃). After comparison, the most efficient medium for shoot regeneration was 1/2 Murashige and Skoog (MS) medium containing 0.5 mg L^–1 BA, 0.05 mg L^–1 NAA and 0.2 mg L^–1 GA₃, which resulted in an average number of induced shoots per explant and shoot length of 5.2 and 1.62 cm at 28 d, respectively. Then, elongated adventitious shoots were transferred to induce roots. 86.7% of shoots was able to root on 1/2 MS medium supplemented with 0.5 mg L^–1 NAA and 0.1 mg L^–1 BA. The earliest rooting time observed was after 21 d and the average root length was up to 3.3 cm after 28 d. Our study shows that C. japonicum can be successfully regenerated through de novo organogenesis, which lays a foundation for future transformation research on this tree.     

桂林岩溶石山密花树群落主要物种的种间关联及群落稳定性

研究群落内主要物种的种间关联及群落稳定性,有助于正确地认识种间关系和物种共存机制,进而能为植被的恢复与重建以及生物多样性保护等提供理论依据。然而桂林岩溶石山密花树群落主要物种的种间关联及群落稳定性仍不明确。基于群落调查数据,采用方差比率法(VR)、χ²检验、Pearson相关检验、Spearman秩相关检验和M.Godron稳定性等方法,对该群落主要物种的种间关联及群落稳定性进行研究。结果表明:(1)该群落乔木层、灌木层和草本层主要物种间的总体关联性均呈显著负关联;(2)χ²检验结果显示,各层次绝大部分种对均无显著关联,种间关联较松散,各物种间呈独立分布格局;(3) Pearson相关检验和Spearman秩相关检验发现,各层次绝大部分种对均呈不显著相关,种间相关性较弱;(4) M.Godron稳定性结果表明,整个群落及其各垂直层次的稳定性测定结果均远离20/80,皆处于不稳定状态。研究显示桂林岩溶石山密花树群落种间关联较松散,各物种呈相对独立分布格局,群落处于不稳定的演替早期阶段。因此,在岩溶地区植被恢复与重建过程中,应将资源利用方式、生物学特征和生态学习性相近,对生境要求相似的优势乡土物种搭配在一起。此外,可采取适当的抚育措施,调节种间关系,提高群落稳定性。