猪伪狂犬病病毒GB株蛋白激酶(PK)基因的克隆及序列分析比较

以伪狂犬病病毒GB株细胞培养物为模板，用PCR方法扩增蛋白激酶(PK)基因，对其进行了克隆、序列测定，并与PRV NIA-3株、Ka株和其他α-疱疹病毒进行了同源性比较。结果显示：所扩增的PK基因片段长1396bp，该片段包含一个开放阅读框(ORF)，长1005bp，编码由334个氨基酸组成的蛋白质。与NIA-3株及Ka株的核苷酸同源性为98．4％和97．5％，基因编码区氨基酸序列的同源性为98．2%和96．4％。具有真核细胞丝氨酸／苏氨酸蛋白激酶氨基酸亚结构域特征序列，而且α-疱疹病毒PK基因具有一些特征性的、共有的氨基酸序列。为今后研究PK基因的功能和构建PK缺失株奠定了基础。     

基于产量阶段的河南省夏玉米灰色——马尔柯夫预测模型

针对较长产量序列纵向及横向可比性较差的问题 ,提出用划分产量阶段的方法进行分段处理。用 GM(1,1)灰色模型分段模拟预测河南省夏玉米单产 ,根据相对气象产量划分产量状态 ,再用马尔柯夫模型转移概率预测系统未来发展方向并修正前者的预报结果。通过模拟和预报 ,准确率都有所提高     

西农萨能羊骨桥蛋白基因OPN的筛选、克隆和生物信息学分析

摘　要: 采用抑制性削减杂交技术构建西农萨能羊乳腺组织在泌乳初期和泌乳高峰期的差异表达基因文库, 筛选泌乳初期和高峰期乳腺组织差异表达基因，通过RT-PCR方法克隆西农萨能羊乳腺组织OPN的CDs区序列,用实时定量PCR分析OPN基因的表达水平变化，并进行序列比对和功能预测。结果成功构建了西农萨能羊不同泌乳期乳腺组织中差异表达基因文库，克隆了乳腺组织OPN基因， GenBank登录号为: EU295699, 开放读码框由834个碱基组成，编码277个氨基酸，前16个氨基酸为推定的信号肽区域。西农萨能羊OPN与绵羊(AF_152416)、牛(NM_174187)、人(DQ_846871)、小鼠(NM_009263) 相应基因核苷酸同源性分别为98%、95%、71%和70%，氨基酸同源性分别为：98%、91%、55%和54%，泌乳初期乳腺组织中mRNA表达水平是泌乳高峰期的5.03倍；预测西农萨能羊OPN较人和小鼠OPN多2个潜在功能基序。     

CrylAb敏感和抗性亚洲玉米螟氨肽酶N基因的克隆及序列差异分析

研究亚洲玉米螟(Ostrinia furnacalis)对Cry1Ab产生抗性的分子机理,对今后制定和实施合理的抗性治理策略具有重要的意义。本研究通过PCR方法克隆和测序,鉴定了Cry1Ab敏感-抗性亚洲玉米螟幼虫中肠氨肽酶N(APN)基因家族的一个成员Ofapn3,其cDNA全序列长3591bp,开放阅读框包括3045bp,编码1014个氨基酸。预测蛋白质分子量为115.1kD,等电点(pI)为4.44。其推导的氨基酸序列中具有鳞翅目昆虫氨肽酶典型结构特征,即N-末端具有18个氨基酸的剪切信号肽,谷氨酸锌化氨肽酶保守结构GAMEN,锌结合位点HEXXHX18E,C-末端具有22个氨基酸的糖基磷脂酰肌醇(GPI)锚信号肽。系统分类归为第3支系。与欧洲玉米螟(Ostrinia nubilalis)的Onapn3(GenBank登录号:ABL01483)同源性为96.6%。与Cry1Ab敏感亚洲玉米螟cDNA相比,抗性品系的开放阅读框中有40个碱基发生了点突变,导致氨基酸序列中有10个氨基酸改变。其中Ser735在抗性品系中突变为Pro的现象在抗性棉铃虫APN3的氨基酸突变中也被检测到。鉴定的Cry1Ab敏感和抗...     

基于图像和光谱信息融合的红茶萎凋程度量化判别

为了实现对红茶萎凋程度量化判别,该研究提出了一种将图像和光谱信息融合后分别与线性判别分析法和偏最小二乘法结合的技术,进行工夫红茶萎凋程度定性判别及儿茶素与氨基酸比值定量预测研究。通过对图像进行主成分分析,筛选出5个特征波长和对应的光谱特征值,基于灰度共生矩阵提取5个特征波长图像的纹理特征值,并采用连续投影算法优选出14个纹理特征值,然后分别以光谱和纹理特征值融合数据建立红茶萎凋程度的线性判别模型和儿茶素与氨基酸比值的偏最小二乘预测模型。结果表明:采用所研究的方法和建立的模型对工夫红茶萎凋程度判别准确率达到94.64%,儿茶素与氨基酸比值预测相关系数为0.8765,预测均方根误差为0.434,预测结果较好。证明应用这两种方法能实现对红茶萎凋程度量化判别。     

流感病毒神经氨酸酶抑制剂的定量构效关系(QSAR)的研究

以31个分子结构与生物活性(-lgIC50)已知的流感病毒神经氨酸酶抑制剂为训练集,选用16个分子的理化描述子,包括分子的形状,静电作用能和范德华作用能以及疏水描述子,用二维定量构效关系(2D-QSAR)方法建立了预测模型。计算中使用偏最小二乘(PLS)和主成分分析(PCA)降低变量维数,减少噪音干扰。用交叉验证法(leave one out,LOO)检验了预测模型的可信性,并用一个由4个抑制剂组成的测试集进一步验证预测模型的预测能力,证实了QSAR预测模型有较好的预测能力。神经氨酸酶抑制剂的QSAR预测模型的建立为抗流感药物的设计和改进提供了有用的研究工具。     

采用改进YOLOv4算法的大豆单株豆荚数检测方法

大豆单株豆荚数检测是考种的重要环节,传统方法通过人工目测的方式获取豆荚类型和数量,该方法费时费力且误差较大。该研究利用大豆单株表型测量仪采集到的表型数据,通过融合K-means聚类算法与改进的注意力机制模块,对YOLOv4目标检测算法进行了改进,使用迁移学习预训练,获取最优模型对测试集进行预测。试验结果表明,该研究模型的平均准确率为80.55%,数据扩充后准确率达到了84.37%,比育种专家目测准确率提高了0.37个百分点,若不考虑5粒荚,该研究模型的平均准确率为95.92%,比YOLOv4模型提高了10.57个百分点,具有更强的检测性能。在简单背景的摆盘豆荚检测中,该研究模型预测的平均准确率达到了99.1%,比YOLOv4模型提高了1.81个百分点,研究结果表明该模型在不同场景下的大豆豆荚检测中具有较强的泛化能力,可为大豆人工智能育种提供参考。     

基于高光谱成像的寒地水稻叶瘟病与缺氮识别

为进行水稻叶瘟病与养分缺失的区分、实现叶瘟病及时、准确的诊断,以大田试验为基础,利用高光谱成像仪获取2个品种的健康、缺氮、轻度感病和重度感病共4类水稻叶片的反射率光谱,对其光谱特性进行分析,并采用多种预处理方法、分别结合偏最小二乘判别分析(partial least squares-discriminate analysis,PLS-DA)和主成分加支持向量机(principle component analysis-support vector machine,PCA-SVM)方法构建水稻叶瘟病识别模型。试验结果显示6个判别模型都获得了较高的识别准确率,经标准正态变量(standard normal variate,SNV)变换预处理的PLS-DA模型获得了最佳的识别结果,预测准确率达100%,经多元散射校正(multiplicative scatter correction,MSC)预处理的PCA-SVM模型的预测准确率也达到97.5%。本研究为水稻叶瘟病的判别和分级提供了新方法,也为稻瘟病大范围遥感监测提供了基础。     

多分类支持向量机在泥石流危险性区划中的应用

以凉山州安宁河流域129个乡镇的泥石流危险性区划资料为依据,随机选取总样本数的2/3和1/2作为训练样本,建立不同数量训练样本下安宁河流域泥石流危险性区划的多分类SVM模型,进行以乡镇为单元的区域泥石流危险性评价研究。评价结果表明,SVM模型的预测精度随着训练样本数量的增加而提高;2个SVM模型对测试样本的预测准确率均高于相应的BP神经网络模型,对训练样本的回判准确率高于或接近于BP神经网络模型。因此,支持向量机方法是一种比神经网络方法具有更优精度和更强泛化性能的新机器学习方法,在泥石流危险性评价实践中具有十分广阔的应用前景和推广应用价值。     

重组伪狂犬病毒(rPRV-GP5)对伪狂犬病最小免疫剂量的测定

伪狂犬病(Pseudorab ies PR)是由伪狂犬病毒(PRV)引起的,可导致多种家畜和野生动物以发热、奇痒(猪除外)及脑髓炎为主要症状的急性传染病。猪是该病毒的中间宿主和传染源,感染后可引起妊娠母猪发生严重的繁殖障碍造成死胎、流产和木乃伊胎,患病仔猪表现为神经症状和呼吸道病,多以死亡告终。在规模化、集约化、现代化高密度养猪地区,PRV可在猪群间不断传播,是危害我国养猪业的一个重要传染病。用疫苗进行免疫接种是预防、甚至消灭伪狂犬病的根本措施。目前伪狂犬病疫苗在我国应用最多的是Bartha-K 61株,该疫苗是20世纪60年代匈牙利Bartha等人将野毒株用鸡胚成纤维细胞反复传代而获得的一个人工致弱疫苗株,该毒株呈现gE基-因缺失表型,其毒力大大减弱,免疫原性很好,是世界上公认的优良疫苗株。我们以Bartha-K61株为载体构建了表达猪繁殖与呼吸综合征病毒(PRRSV)CH-1a株GP5基因的重组伪狂犬病病毒(rPRV-GP5),但GP5基因的插入是否会改变伪狂犬病病毒Bartha-K61株对伪狂犬病的免疫保护效力?为此本研究通过测定重组伪狂犬病毒(rPRV-GP5)对伪狂犬病的最小免疫剂量,评价rPRV-GP5对伪狂犬病的保护效力。24只5~6月龄PRV阴性健康绵羊随机分成6组,每组4只,其中1组为生理盐水对照组,其余5组分别后腿肌注重组伪狂犬病毒(rPRV-GP5)冻干疫苗(病毒含量为5×106.0TCID50/mL)5×104.0TCID50/只、5×103.0TCID50/只、5×102.0TCID50/只、5×101.0TCID50/只和5TCID50/只,免疫后15 d每只绵羊肌注伪狂犬病毒猪源强毒S株1000 LD50。结果表明:对照组和5TCID50/只组的全部绵羊攻毒后4~5 d均出现典型伪狂犬病症状并死亡,5×101.0TCID50/只组4只绵羊中有2只发病并死亡,其余各组全部保护。结论:重组伪狂犬病毒(rPRV-GP5)对绵羊最小免疫剂量为100 TCID50/只,GP5基因的插入没有影响伪狂犬弱毒疫苗Bartha-K61株的免疫原性。     

Environmentally induced changes in amino acid composition in the grain of durum wheat grown under different water and temperature regimes in a Mediterranean environment

Amino acid composition is an important feature in determining the nutritional value of wheat grain for human and animal diets. Environmental conditions are known to influence protein quantity as well as grain production and, in turn, amino acid composition. In this study, grain yield, protein content, and amino acid composition were determined in 10 durum wheat genotypes under three water and temperature regimes in a Mediterranean environment. The highest value for grain-protein content (15.7%) was found in the warmer and driest environment and the lowest (12.8%) in the irrigated environment. Although amino acid composition showed significant variation for all genotypes, with the exception of arginine and cysteine, major changes in amino acid composition were caused by environmental conditions and in particular by water availability and temperature during the grain-filling period, which significantly altered the duration of grain development. The amino acids with the highest percentage of variation between environments were tyrosine (26.4%), lysine (23.7%), methionine (20.3%), threonine (19.3%), and valine (15.6%), whereas phenylalanine (5.1%), glycine (6.4%), and aspartic acid (6.8%) showed the least variation between environments. Whereas the content of glutamine, phenylalanine, and proline increased with the decrease in grain-filling duration, the remaining amino acids tended to diminish, presumably because high temperature and drought favored the deposition of gliadins (proteins particularly rich in glutamine and proline), to the detriment of albumins and globulins (proteins especially rich in threonine, lysine, methionine, valine, and histidine). Despite the negative correlations found between the percentage of protein and its content in essential amino acids, the results indicate that reductions in lysine per unit of food were not very pronounced (0.32 to 0.29 g/100 g of flour) with increases of up 22.7% in grain-protein content, whereas threonine did not change and valine even slightly increased.     

Influence of Malting on the Protein Composition of Spelt (Triticum spelta L.) ‘Frankenkorn’

Although spelt has promising aptitudes for beer and beverage production, almost no investigation about its protein modifications during malting has been conducted. Spelt proteins during the malting process were separated and analyzed, first, according to the isoelectric point and molecular weight (MW) and, second, according to solubility and MW. Moreover, the composition of the free amino acid was determined. Spelt proteins could be divided into a total of 13 zones according to their isoelectric point and MW. Most of the present proteins (90%) had a MW between 35,000 and 60,000 and an isoelectric point between 5.84 and 9.52. The most prominent protein fraction was defined by an isoelectric point between 5.84 and 6.89 and a MW between 35,000 and 55,000. Generally, a protein degradation process during malting was found. The Osborne fractionation revealed an albumin band pattern between 14,000 and 131,700, globulin ranging from 13,400 to 130,500, gliadin from 14,500 to 220,800, and glutenin from 19,800 to 215,300. The free amino acid composition increased in concentration except for asparagine, which decreased during malting. This paper contributes to the understanding of the protein modifications and metabolic changes during the malting process of spelt and facilitates the determination of the potential of this cereal for the production of cereal‐based products.     

Enzyme-linked immunosorbent assay (ELISA) for detection of sulfur-rich protein (SRP) in soybeans (Glycine max L.) and certain other edible plant seeds

As a result of methionine deficiency, legume proteins are considered to be incomplete, and therefore there is a need to explore ways to improve legume protein amino acid balance. Using rabbit anti-soybean sulfur-rich protein (SRP) polyclonal antibodies (pAb), sensitive immunoassays (nanogram sensitivity) were developed. The immunoassays detected SRP in all soybean seeds and soybean-based commercial samples examined. In addition, the presence of pAb cross-reactive proteins was detected in certain dry beans and oilseeds. The cross-reactive proteins were isolated using purified IgG-based immunoaffinity column chromatography. Biochemical analyses including N-terminal amino acid sequencing and amino acid composition indicated that the cross-reactive proteins were comparable to soybean SRP. The cross-reactive proteins contained methionine (1.6-2.4 residues/100 residues) and cysteine (2.4-3.6 residues/100 residues), which satisfies the FAO/WHO recommended pattern for sulfur amino acids in both adults and children (2-5 years old). The results suggest the presence of constitutive SRPs in several dry beans and oilseeds.     

Modeling protein functional properties from amino acid composition

Physicochemical and functional properties of proteins were modeled as a function of the contributions of each of the 20 coded amino acids to three (z-scores) or five (extended z-scores) amino acid principal properties using partial least squares regression. The five term models were in all cases stronger in both fit and prediction than the three term models, indicating that useful information is contained in the fourth and fifth property scores. Models predicting protein hydrophobicity (R = 0.932), viscosity (R = 0.737), and foam capacity (R = 0.880) from amino acid composition rather than sequence were obtained. It is likely that additional functional and physicochemical properties of proteins can be modeled in this way.     

High molecular weight melanoidins from coffee brew

The composition of high molecular weight (HMw) coffee melanoidin populations, obtained after ethanol precipitation, was studied. The specific extinction coefficient (K(mix)) at 280, 325, 405 nm, sugar composition, phenolic group content, nitrogen content, amino acid composition, and non-protein nitrogen (NPN) content were investigated. Results show that most HMw coffee melanoidins are soluble at high ethanol concentrations. The amino acid composition of the HMw fractions was similar, while 17% (w/w) of the nitrogen was NPN, probably originating from degraded amino acids/proteins and now part of melanoidins. A strong correlation between the melanoidin content, the NPN, and protein content was found. It was concluded that proteins are incorporated into the melanoidins and that the degree of chemical modification, for example, by phenolic groups, determines the solubility of melanoidins in ethanol. Although the existence of covalent interaction between melanoidins and polysaccharides were not proven in this study, the findings suggest that especially arabinogalactan is likely involved in melanoidin formation. Finally, phenolic groups were present in the HMw fraction of coffee, and a correlation was found with the melanoidin concentration.     

Nutritional composition of Sanchi (Panax notoginseng) seed and its potential for industrial use

Sanchi (Panax notoginseng) root has been widely used as traditional herb to stanch blood, disperse gore and reduce pain in China for centuries. However, the nutritional information about its seed was unknown. Sanchi seeds obtained from Yunnan Province were analyzed for their nutritional composition, fatty acid, amino acid profile and sugar contents. Sanchi seeds were abundant in fats (46.35 %) and proteins (23.90 %). Notably, seven fatty acid compositions were determined and abundant in unsaturated fatty acid (99.56 %), containing 95.71 % oleic acid. Sanchi seed proteins were rich in glutenin, globulin and albumin (28.63, 27.83 and 26.81 %, respectively). Sanchi seed contain 17 kinds of amino acids, of which nine were essential amino acids, accounting for 41.30 %. These nutritional compositions indicate that Sanchi seed has the potentiality to be exploited as human edible oil, industrial use, new medicine or healthcare products for diabetes patients due to low sugar content.     

Characterization of royal jelly proteins in both Africanized and European honeybees (Apis mellifera) by two-dimensional gel electrophoresis

In this study, the proteins contained in royal jelly (RJ) produced by Africanized honeybees and European honeybees (Apis mellifera) haven been analyzed in detail and compared using two-dimensional gel electrophoresis, and the N-terminal amino acid sequence of each spot has been determined. Most spots were assigned to major royal jelly proteins (MRJPs). Remarkable differences were found in the heterogeneity of the MRJPs, in particular MRJP3, in terms of molecular weights and isoelectric points between the two species of RJ. Furthermore, during the determination of the N-terminal amino acid sequence of each spot, for the first time, MRJP4 protein has been identified, the existence of which had been only implied by cloning of its cDNA sequence. The presence of heterogeneous bands of glucose oxidase was also identified. Thus, the results suggest that two-dimensional gel electrophoresis provides a suitable method for the qualitative analysis of the proteins contained in RJ derived from different honeybee species.     

Relationship between varietal amino acid profile of grapes and wine aromatic composition. Experiments with model solutions and chemometric study

Synthetic solutions containing amino acids, sugar, water, and yeast nutrients have been fermented by Saccharomyces cerevisiae, and the volatile composition of the fermented media has been analyzed by GC. Eleven amino acid compositions imitating the characteristic amino acid profile of 11 different grape varieties were tested. Significant differences in the levels of some important volatile compounds (ethanol, ethyl acetate, acetic acid, higher alcohols and some of their acetates, methionol, isobutyric acid, ethyl butyrate, and hexanoic and octanoic acids) were found. The levels of some of the volatile compounds are well correlated with the aromatic composition of wines made with grapes of the same varieties. A multiple linear regression study produced good models for most of the odorants for which the level was related to the must amino acid composition. Partial least-squares regression models confirm that amino acid composition explains a high proportion of the variance in the volatile composition and show that the relationship between both sets of variables is highly multivariate. According to the different models, the levels of some byproducts of fatty acid synthesis are related to threonine and serine, the level of beta-phenyletanol is closely related to the level of phenylalanine, and methionol is strongly correlated to the must methionine contents. The addition of selected amino acids to different musts confirms the previous observations.     

Altered amino acid profile of arbuscular mycorrhizal maize plants under low temperature stress

We investigated the effect of arbuscular mycorrhiza (AM) on amino acid concentration and composition of maize plants under low‐temperature stress. The AM plants had higher amino acid concentrations than the non‐AM pants. The concentrations of Thr, Lys, Gly, Ala, His, and Ile of the AM plants were higher than non‐AM plants. The results show that low‐temperature stress decreased the concentrations of amino acids and altered their composition.     

Physicochemical and functional properties of hemp (Cannabis sativa L.) protein isolate

The amino acid composition and physicochemical and functional properties of hemp (Cannabis sativa L.) protein isolate (HPI) were evaluated and compared with those of soy protein isolate (SPI). Edestin, a kind of hexameric legumin, was the major protein component. HPI had similar or higher levels of essential amino acids (except lysine), in comparison to those amino acids of SPI. The essential amino acids in HPI (except lysine and sulfur-containing amino acids) are sufficient for the FAO/WHO suggested requirements for 2-5 year old children. The protein solubility (PS) of HPI was lower than that of SPI at pH less than 8.0 but similar at above pH 8.0. HPI contained much higher free sulfhydryl (SH) content than SPI. Differential scanning calorimetry analysis showed that HPI had only one endothermic peak with denaturation temperature (T(d)) of about 95.0 degrees C, attributed to the edestin component. The T(d) of the endotherm was nearly unaffected by 20-40 mM sodium dodecyl sulfate but significantly decreased by 20 mM dithiothreitol (P < 0.05). The emulsifying activity index, emulsion stability index, and water-holding capacity of HPI were much lower than those of SPI, and the fat adsorption capacity was similar. The data suggest that HPI can be used as a valuable source of nutrition for infants and children but has poor functional properties when compared with SPI. The poor functional properties of HPI have been largely attributed to the formation of covalent disulfide bonds between individual proteins and subsequent aggregation at neutral or acidic pH, due to its high free sulfhydryl content from sulfur-containing amino acids.