新疆梨PsSFBB基因克隆及其生物信息学分析

[目的]对新疆梨PsSFBB基因进行克隆,并对其进行生物信息学分析。[方法]以新疆梨品种棋盘梨的花药为材料,利用RT-PCR和RACE技术对其PsSFBB基因进行克隆,并对克隆出的基因进行生物信息学分析。[结果]试验克隆到了一个全长为1 231 bp的PsS-FBB-α基因(Genbank登录号为EU909687),命名为PsSFBB6-α。该基因编码378个氨基酸,在N末端含有一个约50个氨基酸组成的F-box基序。经生物信息学分析,推测PsSFBB6-α蛋白的分子式为C2000H3034N517O558S23,相对分子质量为44 987.5,等电点为6.02,二级结构以α螺旋为主;理论推导半衰期大约为30 h,不稳定参数为55.21,属于不稳定蛋白;并且推测此蛋白为亲水性、非分泌型蛋白,在基质内起裂合酶的作用,特异性的识别底物,正好吻合了F-box蛋白的作用。[结论]为进一步研究SFBB蛋白和自交不亲和机理奠定了基础,也为新疆梨自交亲和品种的选育和生产中科学配置授粉树以提高产量和品质提供理论了依据。     

魏氏梭菌α毒素快速诊断金标试纸条的研制

【目的】研制魏氏梭菌α毒素快速诊断金标试纸条,为魏氏梭菌病的快速诊断提供新方法。【方法】采用分子生物学方法制备魏氏梭菌α毒素,将其免疫家兔制备多克隆抗体。用制备的抗魏氏梭菌α毒素多克隆抗体作为胶体金标记抗体和检测线上的捕获抗体,制备魏氏梭菌α毒素快速诊断金标试纸条,对其特异性、敏感性、稳定性和重复性进行检测。用制备的魏氏梭菌α毒素快速诊断金标试纸条对临床样品进行检测,比较其检测结果与PCR检测结果的符合率。【结果】成功制备了魏氏梭菌α毒素多克隆抗体。成功研制了α毒素快速诊断金标试纸条,其检测线上抗体的最适包被质量浓度为0.06 μg/mL。研制的试纸条仅与魏氏梭菌α毒素发生反应,且不同批次的试纸条重复检测结果无差异,表明试纸条具有较高的特异性和重复性;该试纸条最低可检测1.40 mg/mL的毒素量,与PCR方法测得结果符合率为80.77%,基本能满足实际应用要求。【结论】成功研制了魏氏梭菌α毒素快速诊断金标试纸条,该试纸条具有快速、特异、敏感、稳定等优点,适用于对魏氏梭菌病进行普查和检疫,具有良好的应用前景。     

Assessing the severity of canine pancreatitis

The objective of this study was to determine whether laboratory testing currently available is able to provide prognostic information in canine pancreatitis. A prospective study of dogs with naturally occurring pancreatitis was undertaken. Twenty-two cases with histologically confirmed pancreatic inflammation were included in the study. Each dog had routine haematology parameters, serum biochemistry (including lipase and amylase), serum trypsin-like immunoreactivity and trypsinogen activation peptides (TAP) in urine and plasma measured. Twelve of the dogs were classified as having severe disease. These dogs had statistically significant increases in urinary TAP-creatinine ratio (UTCR) measurement, serum lipase, serum phosphate and serum creatinine concentrations. Additionally dogs with severe pancreatitis had significantly decreased urine specific gravity levels. The most sensitive and specific test to assess the severity of pancreatitis was the measurement of UTCR.     

白唇鹿血清同工酶酶谱的电泳研究

采用聚丙烯酰胺凝胶电泳地对３４头白唇夜以继日的４种血清同工酶酶谱及其多态性进行研究。结果（１）酯酶包括ＥＳ１，ＥＳ２和ＥＳ３三种同工酶，其中ＥＳ２存在多态性；（２）碱性磷酸酶存在在ＡＬＰＢ和ＡＬＰＯ两种表型，以ＡＬＰＯ型为优势表型（６７．６５％）；（３）淀粉酶包括ＡＭＹ１，ＡＭＹ２和ＡＭＹ３三种同工酶，ＡＭＹ１和ＭＡＹ２两种同工酶存在多态性，（４）白唇鹿的乳酸脱氢酶显现出与众不同的同工酶酶谱。     

Dynamics of Environmental Humidity and Water Content in Tobacco Leaf and Metabolism of Starch During Curing

Effect of environmental humidity and water content in tobacco leaf on starch metabolism was studied by using the electric-heated auto-controlled flue-curing barn supplied by Henan Agricultural University, China. The results indicated that the degradation of starch and decrease of water content in tobacco leaf during early yellowing at low humidity was the most rapid, and the water loss was the highest while the lowest under high humidity. The duration for starch degradation under low humidity was longer than that of CK. So the starch residue in cured leaf of different treatment took the order of high humidity yellowing treatment＞low humidity yellowing treatment＞ CK. When the leaf water content was decreased to around 50%, the starch degradation became slow and the content of starch was stable. Starch degradation and decrease of leaf water content was not synchronous. Starch in tobacco leaf during yellowing degraded more rapidly when humidity was decreased at a high speed, but the degradation stopped earlier at late stage. There was a quicker and higher degradation of starch under high environmental humidity. When the humidity decreased to 70 %,the content of starch was stable. The activity of amylase began to decrease when relative humidity was below 75 %, but it kept a high level of activity when the environmental humidity was below 70 %.     

大豆猝死综合症病原菌北美种的PCR鉴定

 应用等位基因特异性PCR(allele-specific PCR,AS-PCR)技术,建立了大豆猝死综合症(sudden death syndrome of soybean,SDS)病原菌北美种Fusarium virguliforme与其近似种Fusarium phaseoli(菜豆根腐病菌)的鉴别方法。针对翻译延长因子(EF-1α)基因上的3个SNP(单核苷酸多态性)位点,参照引物设计原则和等位基因特异引物(allele-specific primer,ASP)的要求设计了1对引物Fsg-α-1/2,能特异地扩增F.virguliforme,产生327 bp的电泳条带。另外,根据ITS区序列设计了1对通用引物Fu1/2,能扩增F.virguliforme和F.phaseoli,产生452 bp的电泳条带。本实验在传统的AS-PCR基础上进行改进,引入Fu1/2作为阳性质控引物,将ASP的特异性与双重PCR的严谨性相结合,建立了简便可靠的SDS病原菌鉴定方法。     

壳聚糖包衣对杂交水稻种子发芽和幼苗耐盐性的影响

用1.5%壳聚糖和1.5%壳聚糖+10 mmol/L脯氨酸(Proline)包衣处理两个杂交稻品种"汕优10"和"二优培九"种子, 包衣和未包衣种子分别在H2O、 50 mmol/L、 100 mmol/L和150 mmol/L的NaCl溶液中在30℃下发芽, 并测定种子发芽率、淀粉酶活性、幼苗耐盐指数、耐盐比率及幼苗脯氨酸、可溶性糖、果糖和蔗糖等相容性溶质含量. 结果表明,     

酶水解法制取大米蛋白的机理研究

(1郑州轻工业学院食品与生物工程学院,郑州 450002;2河南科技大学食品与生物工程学院,洛阳 471003; 3江南大学食品学院,无锡 214036)     

运用农杆菌介导法将PTA和反义Wx双基因导入水稻的研究

利用根癌农杆菌将含多基因(hyg选择标记基因、反义Wx 基因、PTA基因、GFP和GUS报告基因)的pCAMBIA1304载体导入水稻品种(501R、中花9号和日本晴)的幼胚愈伤组织,分别在含35mg/L、45mg/L和65mg/L潮霉素浓度的筛选培养基上筛选获得抗性愈伤。后经PCR检测,从415株T0代再生植株中选出92株(其中501R、中花9号和日本晴分别为4株、43株和45株)。对这些转基因后代植株进行Southern blotting分析表明：hpt、反义Wx 和 PTA基因已经整合进植物基因组中,绝大多数转基因植株后代的表型正常。成熟种子直链淀粉含量分析表明,部分转基因植株的T1代种子中的直链淀粉含量有部分程度的下降,最低的已下降至6.3%,与对照相比下降了9.8%。     

华南5号和华南6号木薯体胚发生 过程中酯酶和淀粉酶同工酶分析

以木薯华南5号(SC5)和华南6号(SC6)两个品种的成熟体胚子叶为外植体进行体胚诱导培养。对诱导培养0、5、10、20、30 d的材料,采用聚丙烯酰胺垂直板凝胶电泳比较其酯酶(EST)和淀粉酶同工酶酶谱的差异。结果表明,酯酶同工酶的表达在品种间存在差异。SC5中E2和E3条带与SC6中E’2和E’3条带迁移率不一致,可能是品种差异的原因。表达最强的酶带在SC5中是E3,SC6是E’4。两个品种淀粉酶同工酶的区别在于,SC5诱导过程中最多时有3条酶带出现,而SC6自始至终只有1条酶带出现。SC5酶带最亮时出现在体胚诱导培养5 d时,且此时也是酶带最多的时候。SC6酶带出现的最亮时是在外植体诱导前和诱导培养20 d时。