Oxygen diffusion-concentration in erythrocyte plasma membranes studied by the fluorescence quenching of anionic and cationic pyrene derivatives

Fluorescence quenching by oxygen of cationic [pyrene-(CH₂) _n N(CH₃) ₃ ⁺ ;n=1, 4, and 11] and anionic [pyrene-(CH₂) _n CO ₂ ^– ,n=3, 8, 11, and 15] probes was investigated in erythrocyte plasma membranes (leaky) in order to assess the ability of oxygen molecules to interact with solutes located at different positions in the membrane. The pseudounimolecular quenching rate constants measured increase, both for cationic and anionic probes, whenn increases. These results are interpreted in terms of an increased oxygen solubility toward the center of the membrane interior, and imply that lateral diffusion contributes more than transverse diffusion to total oxygen mobility. For all of the probes considered, quenching rates increase whenn-alkanols are added. The effect observed increases whenn decreases and when the size of then-alkanol alkyl chain increases. Arrhenius-type plots for the quenching rate constants show noticeable downward curvatures. Average (0–40°C) activation energies are 6 kcal/mol.Abbreviations EPM erythrocyte plasma membrane - PMTMA (1-pyrenyl)methyltrimethyl-ammonium - PBTMA 4-(1-pyrenyl)butyltrimethylammonium - PUTMA 11-(1-pyrenyl)-undecyltrimethylammonium - PB 4-(1-pyrenyl)butanoate - PN 9-(1-pyrenyl)nonanoate - PD 12-(1-pyrenyl)dodecanoate - PHD 16-(1-pyrenyl)hexadecnoate     

The novel avirulence effector AlAvr1 from Ascochyta lentis mediates host cultivar specificity of ascochyta blight in lentil

Ascochyta lentis is a fungal pathogen that causes ascochyta blight in the important grain legume species lentil, but little is known about the molecular mechanism of disease or host specificity. We employed a map‐based cloning approach using a biparental A. lentis population to clone the gene AlAvr1‐1 that encodes avirulence towards the lentil cultivar PBA Hurricane XT. The mapping population was produced by mating A. lentis isolate P94‐24, which is pathogenic on the cultivar Nipper and avirulent towards Hurricane, and the isolate AlKewell, which is pathogenic towards Hurricane but not Nipper. Using agroinfiltration, we found that AlAvr1‐1 from the isolate P94‐24 causes necrosis in Hurricane but not in Nipper. The homologous corresponding gene in AlKewell, AlAvr1‐2, encodes a protein with amino acid variation at 23 sites and four of these sites have been positively selected in the P94‐24 branch of the phylogeny. Loss of AlAvr1‐1 in a gene knockout experiment produced a P94‐24 mutant strain that is virulent on Hurricane. Deletion of AlAvr1‐2 in AlKewell led to reduced pathogenicity on Hurricane, suggesting that the gene may contribute to disease in Hurricane. Deletion of AlAvr1‐2 did not affect virulence for Nipper and AlAvr1‐2 is therefore not an avirulence gene for Nipper. We conclude that the hemibiotrophic pathogen A. lentis has an avirulence effector, AlAvr1‐1, that triggers a hypersensitive resistance response in Hurricane. This is the first avirulence gene to be characterized in a legume pathogen from the Pleosporales and may help progress research on other damaging Ascochyta pathogens.     

棉蚜抗药性机理研究进展

棉蚜的抗药性涉及到代谢酶活力的提高、靶标部位的不敏感以及表皮穿透作用的降低。该文重点概述了棉蚜代谢抗性和靶标抗性方面的研究进展 ;介绍了棉蚜抗性中存在的交互抗性、负交互抗性 ,讨论了抗性稳定性以及寄主植物和转基因棉对棉蚜抗药性的影响。     

萃取-结晶法制备辣椒碱类化合物工艺研究

为开发一条从辣椒精制备辣椒碱类化合物的工艺路线,考察了皂化、萃取和结晶条件对辣椒碱类化合物收率和纯度的影响,获得了纯化辣椒碱类化合物的最佳工艺条件:5%氢氧化钠溶液皂化辣椒精,皂化液酸化后用乙醚萃取2次,乙醚相用氢氧化钠水溶液萃取3次,所得氢氧化钠萃取液调pH值后结晶,得到辣椒碱类化合物,纯度为98.73%,总收率为78.89%.     

江西省主要地方鸡种的RAPD分析及其群体遗传关系的研究

采用RAPD技术分析了南城黑鸡、余干五黑鸡、泰和乌骨鸡、东乡绿壳蛋鸡、崇仁麻鸡、宁都三黄鸡、广丰白耳黄鸡和万载康乐黄鸡等8个江西地方鸡种、1个培育鸡种--景德黄鸡以及1个外来鸡种--以色列隐性白羽鸡基因组池DNA的多态性,经OPERONA,B,C,D,G5组100种随机引物扩增筛选,14种引物获得了33个多态标记.各品种间的遗传距离指数的计算结果和UPGMA聚类关系图表明:在10个鸡种间,南城黑鸡和余干五黑鸡的遗传距离最近,两者有着极密切的亲缘关系.据此认为这两个鸡群可视为同一品种的两个地方系.此外,广丰白耳黄鸡和景德黄鸡也表现出较密切的亲缘关系,这与景德黄鸡在培育过程中曾掺有白耳黄鸡血缘的选育历史相吻合。 Abstract:Random amphfied polymorphic DNAs (RAPD) analysis was explored to investigate genetic polymorphisms of pooled genomic DNA from 8 native and 1 cultivated chicken breeds in Jiangxi province as well as 1 exotic chicken breed,which include Guangfeng baier huang,Jingde huang,Yugan wuhei,Nancheng hei,Congren ma,Taihe Silky Fowl,Ningdu san huang,Dongxiang green-shell,Wanzai kangle huang and Israel Recessive White Feather Fowl.of 100 OPERON random primers screened,14 primers produced 34 polymorphic RAPD markers.The genetic distance index and UPGMA dendrogram indicated that Nancheng hei and Yugan wuhei chickens had very closely genetic relationship,the genetic distance between the two populations was closer than any other pair of breeds.Therefore,the two chicken populations were suggested to be classified into one breed.In addition,Guangfeng baier huang and Jingde huang shown intimately relationship,it was consistent with the cultivation history of Jingde huang chicken breed,in which Guangfeng baier huang was introduced as one of parental breed.     

赣南地区农业生态经济系统耦合态势的时空演变

赣南地区地处南方丘陵山地区域,是重要的生态功能区,其农业经济发展与生态功能保护的关系备受关注。促进农业生态经济系统的耦合协调发展,是赣南地区可持续发展的重要内容。利用2000-2015年赣州市各县（市）统计年鉴数据及遥感数据,构建了基于耦合协调度模型的区域农业生态经济系统的评价指标体系,开展时间和空间尺度上的农业生态经济系统的耦合协调发展研究。结果表明：（1）在时间尺度上,农业生态系统变化不大,在空间尺度上县（市）之间因自然条件的不同略有差异;农业经济综合评价值在2000-2015年之间有较大的增长,且不同县（市）间存在差异,农业经济系统的相对滞后是限制农业生态经济系统不能协同发展的原因。（2）耦合度和耦合协调度随时间而增长,大部分县（市）的耦合度经历了从中度耦合、高度耦合、再到极度耦合的过程,其中极度耦合的占比从2000年的0%提升到2015年的94%;耦合协调度从基本、中度协调耦合到中度、高度协调耦合,且都呈现以赣县、于都县、兴国县为主的中部较低,而南北部较高的态势。（3）2010年前,大部分县（市）的农业生态经济系统发展类型长期处于农业经济极度滞后状态,随后逐步向农业经济严重滞后过渡。农业生态经济系统耦合态势的分析有助于明晰当地的农业生态经济系统的发展、演变规律,促进农业系统的协调发展。     

基于CRISPR/Cas9系统对干酪乳杆菌进行红色荧光蛋白标记

本实验利用CRISPR/Cas9系统对干酪乳杆菌(Lactobacillus casei) LC2W进行红色荧光蛋白(red fluorescent protein,RFP)标记,用于研究干酪乳杆菌在肠道内的分布和定植状况,评价其作为益生菌的功能。首先,基于本实验室已有的干酪乳杆菌CRISPR/Cas9编辑质粒pLCNICK-1628构建重组质粒pLCNICK-1628-RFP,电转入干酪乳杆菌LC2W感受态细胞中,使干酪乳杆菌基因组中的LC2W-1628基因被红色荧光蛋白基因替换,从而使干酪乳杆菌LC2W能表达出红色荧光蛋白。得到红色荧光标记的干酪乳杆菌LC2W突变株后,测定了其荧光强度-OD600标准曲线,发现RFP在干酪乳杆菌LC2W中能稳定表达。     

重组人KGF制备工艺和活性检测方法的建立

目的:在毕赤酵母中实现了KGF的高效表达,并初步建立了生物学活性检测方法.方法:合成5'端缺失69个核苷酸的KGF基因序列,克隆入pPIC9并转化毕赤酵母菌株GS115中,经诱导表达.发酵液上清采用脱盐层析和阳离子交换层析进行分离纯化.利用貂肺上皮细胞(Mv-1-Lu)检测其生物学活性.结果:表达水平达到了110mg/L发酵液;表达产物经一步离子交换层析就能得到有效分离,总收率在50mg/L发酵液以上;纯化的rhKGF生物学活性与KepivanceTM相当.结论:rhKGF制备工艺和检测方法的建立将为该因子的规模化生产和进一步的临床应用提供良好基础.     

hβ2m/HLA-B2704双转基因小鼠的制备及β2m对HLA-B2704表达的影响

制备hβ2m HLA B2 70 4双转基因小鼠 ,研究hβ2m对HLA B2 70 4表达的影响 .通过hβ2m转基因小鼠和HLA B2 70 4转基因小鼠交配 ,出生动物及后代经PCR初步筛选 ,采用Southern杂交对经PCR初步筛选的hβ2m HLA B2 70 4双转基因阳性小鼠基因组DNA标本作进一步鉴定 .阳性者进行RT PCR和流式细胞光度术检测其在mRNA和蛋白水平的表达 .hβ2m阳性小鼠和HLA B2 70 4阳性小鼠交配 ,生仔鼠 6 7只 ,其中 2 8只仔鼠同时整合hβ2m和HLA B2 70 4基因 .Southern杂交证实 ,阳性鼠同时都含有hβ2m和HLA B2 70 4基因 .阳性小鼠的皮肤、结肠、睾丸和脾脏组织中均有HLA B2 70 4和hβ2m基因的mRNA表达 ,其外周血淋巴细胞膜上HLA B2 70 4基因的蛋白表达为 35 87% ,在HLA B2 70 4单转基因小鼠外周血淋巴细胞膜上HLA B2 70 4基因的蛋白表达为 7 87% (Histogramsta tistics) .成功地制备了hβ2m HLA B2 70 4双转基因小鼠 .在hβ2m HLA B2 70 4双转基因小鼠的外周血淋巴细胞膜上 ,HLA B2 70 4基因在蛋白水平表达较高 ,而在HLA B2 70 4单转基因小鼠中则表达不明显 .hβ2m可与HLA B2 70 4重链分子结合 ,稳定和增高其在淋巴细胞膜上的表达