牛分枝杆菌PE_PGRS62基因克隆及在耻垢分枝杆菌中的表达

为构建表达牛分枝杆菌(M.bovis)PE_PGRS62蛋白的重组耻垢分枝杆菌,本研究以M.bovis基因组DNA为模板PCR扩增PE_PGRS62基因,获得大小约为1 515 bp的目的片段,并克隆到大肠杆菌-分枝杆菌穿梭表达载体pMV261中,将重组pMV-PE_PGRS62穿梭表达载体电转化到耻垢分枝杆菌内,42℃热诱导重组耻垢分枝杆菌,通过SDS-PAGE和免疫印迹分析表达产物并鉴定其生物学活性。经SDS-PAGE分析,表达的PE_PGRS62蛋白分子量约为60 ku,免疫印迹结果表明,表达的PE_PGRS62蛋白与兔抗M.bovis阳性血清反应形成一条特异性蛋白条带。本实验为进一步研究M.bovis致病机理奠定了基础。     

甜菜夜蛾核多角体病毒Se62基因的克隆与结构分析

[目的]克隆并分析甜菜夜蛾核多角体病毒（却odoptera exigua muhicapsid nucleopolyhedrovirus,SeMNPV）ORF62全长基因（Se62）。[方法l采用PCR的方法扩增Se62基因,将其克隆至pMDl8．T载体上,获得了重组质粒T．5e62,进行序列测定和分析。[结果]获得大小为1128bp左右的序列,扩增序列与GenBank登录序列（No．NC_002169）核苷酸同源性100％。序列分析表明,Se62基因编码蛋白（SE62）存在明显的跨膜区域,有多个蛋白激酶c、酪蛋白激酶Ⅱ磷酸化位点和N．糖基化位点。蛋白序列比对结果表明,SE62属于杆状病毒P48蛋白超级家族成员,与苜蓿丫纹夜蛾核多角体病毒（Autographa califorica muhicapsid nucleopolyhedrovirus,AcMNPV）P48蛋白的同源性为52％,其在病毒复制中的作用值得关注。[结论]成功克隆出Se62基因,为研究其在病毒复制中的功能奠定了基础。     

The relative contributions of soil hydrophilicity and raindrop impact to soil aggregate breakdown for a series of textured soils

Soil aggregate breakdown is the first key factor that causes soil erosion. At present, research on the mechanisms of soil aggregate breakdown during rainfall is common. However, the research to on quantifying the relative contributions of internal and external forces to aggregate breakdown remains limited. This paper was conducted to analyse the relative contribution of internal and external forces to aggregate disintegration and the factors affecting aggregate stability during rainfall. Soil aggregates with a series of textures were selected as test soil samples; deionized water was employed as the soaking solution and rainfall material in static disintegration experiments and rainfall simulation tests. The effect of internal force (soil hydrophilicity) on aggregate disintegration was analysed by the static disintegration method, and the combined effects of internal force (soil hydrophilicity) and external force (raindrop impact) on soil aggregate breakdown were analysed by rainfall simulation experiments. The results indicated that external force caused more severe soil aggregate breakdown than internal force, and the crushed aggregate was mainly distributed in the range of 2–0.25 mm. With increasing rainfall kinetic energy, the degree of aggregate breakdown increased gradually, and the degree of aggregation of the soil particles decreased gradually. Furthermore, soil aggregates with a high clay content (> 30%) were more stable than medium-clay (20–30%) and low-clay (< 20%) soil aggregates, and the correlation coefficient provided a good representation of the relationship between the clay content and soil aggregate stability index (ASI). Therefore, external force contributed more to soil aggregate breakdown than internal force during rainfall, and clay plays an important role in aggregate stability. The results of this study are of great significance for elaborating the mechanism and factors affecting aggregate breakdown.