种子特异启动子的克隆及植物表达载体构建

[目的]启动子是植物基因工程中一个重要工具,对构建植物生物反应器有着重要意义。8S球蛋白是绿豆中含量最丰富的种子贮藏蛋白,因此,其调控序列可能提供了一个很好来源的种子特异性启动子。[方法]通过基因组步移技术克隆了绿豆8s球蛋白a亚基基因8SGα的启动子区域,基因组步移使用的引物根据绿豆8SGα的cDNA序列设计。[结果]通过3轮PCR的扩增,得到了472bp的上游序列片段,构建了植物双元表达载体pBI－8SGα-GUS。[结论]研究结果可用于转化植物,并于转基因植物中分析启动表达的时空特异性及表达强度,并为植物种子生物反应器提供重要元件。     

Protein electrophoresis of psittacine plasma

BACKGROUND: Although protein electrophoresis (EPH) has been widely applied in human and veterinary medicine, it has only recently been implemented in the analysis of avian samples. OBJECTIVE: The purpose of this study was to examine the application of protein EPH to the analysis of psittacine plasma samples. Our goals were to describe protein fraction mobility, establish reference intervals for some common species, determine the coefficient of variation (CV) of the chosen method, and examine the effects of sample handling and sample condition. METHODS: Heparinized plasma samples from several common psittacine species (minimum sample size 50 each) were examined using the Beckman Paragon system and SPEP-II gels. Total protein was measured by refractometry. Reference intervals (95%) were calculated by the rank methods. RESULTS: Fraction migration patterns were found to vary among common psittacine species. Day-to-day CV for the EPH fractions ranged from 2.2% to 10.5%; within-run CV ranged from 4.8% to 10.8%; and total CV ranged from 3.2% to 14.8%. The highest CV was noted for the poorly defined alpha-globulin fraction. Prolonged refrigeration, repeated freeze-thawing, hemolysis, and lipemia altered the results. CONCLUSIONS: Protein fractions from psittacine species were variable in terms of migration pattern and protein concentration, which necessitates the use of species-specific reference intervals. Avian protein electrophoretic patterns and values should be interpreted based on knowledge of the CV associated with the technique as well as on the effects of sample handling and condition.     

Genetic Diversity of Soybean (Glycine Max (L.) Merrill)7S Globulin Protein Subunits

Out of 851 soybean accessions from Vietnam, China and Japan analyzed for 7S β-subunit variants, a new β-reduced subunit line with normal growth was collected from the Mekong Delta, Vietnam. Protein of the `β-reduced' line is composed of β-reduced and extremely low-β types. (α-null + β-reduced) type and β-null (or extremely low-β) type were screened in the progeny seeds of the Japanese mutant `(α + β) null' line. Therefore, recombination between α- and β-subunits was identified. By comparing the nucleotide sequence of the partial β-subunit gene of Enrei (standard), Mo-shi-dou Gong 503 (α + β low), β-5₃₃ seed (β-reduced), β-5₇ seed (extremely low-β), and (α + β)-null₁₆((α + β) null) seed, we found that the base `T' at 166 bp, in Enrei changed to `G' in Mo-shi-dou Gong 503, β-5₃₃ and β-5₇. Using the (α + β)-null₁₆ individual as template, a distinct 305 bp β-subunit gene fragment was identified, instead of a 285 bp fragment.     

冬小麦籽粒蛋白质组分的形成规律及其调控研究

冬小麦灌浆期间籽粒蛋白质各组分形成时间不同,清蛋白和球蛋白主要在籽粒形成期和灌浆前期形成;醇溶蛋白和谷蛋白主要在灌浆期形成,直至成熟后仍有增长。增加施N量,或中后期追N或喷N等措施对增加醇溶蛋白和谷蛋白效果显著,对籽粒的加工品质有所改善,但对清蛋白和球蛋白含量影响较小,对改善营养品质作用有限。     

Functionality of Succinylated Brazil Nut (<Emphasis Type="Italic">Bertholletia excelsa HBK</Emphasis>) Kernel Globulin

One of the possible ways to improve the utilisation of defatted Brazil nut kernel flour, a by-product of oil extraction industries, is to improve its functional properties by chemical modification as it possesses very modest functional characteristics. Succinylated Brazil nut kernel globulin at 55.8%, 62.4% and 72.0% level showed a positive effect on functionality. The solubility of acylated globulin was improved above pH 4.0 butwas reduced in the pH range of 3.0-4.0. Water absorption (1.96-4.00, 4.12, and 4.21 ml/g protein), oil absorption capacity (1.44-2.72, 2.80 and 2.94 ml/g protein) and apparent viscosity of the succinylated globulin increased with increase in the level of succinylation. The extent of modi-fication also influenced emulsifying capacity, which showed a decrease at pH 3.0, but was increased at pH 5.0, 7.0 and 9.0. Highest emulsion activity (approximately 63.0%) was observed at pH 3.0, followed by pH 9.0 and pH 7.0 and, least (about 11.8%) at pH 5.0. Emulsion stability also followed similar behaviour as that of emulsion activity. The improved functional properties of succinylated Brazil nut kernel globulin could be explored in a variety of food formulations such as high protein drinks, soups, bakery and meat products as well as in salad dressings and mayonnaise as an emulsifier.     

Denaturation by heat,sodium dodecyl sulphate and dithiothreitol of globulins and phaseolin from dry bean (Phaseolus vulgaris L.)

The electrophoretic pattern of the untreated common bean globulin cv. Flor de Mayo had 7 protein fractions ranging from 620 to 120 kilodaltons (kd). The last molecular weight corresponds to the monomeric form. One of the objectives of the present work was to establish a comparison among denaturation by heat, sodium dodecyl sulphate (SDS) and dithiothreitol (DTT). At pH 6.0, two bands were resistant to heat treatment, after SDS treatment of the untreated globulin several bands disappeared and two new bands with 26 and 15 kd appeared. DTT did not change the electrophoretic pattern, due to the small quantity of free SH groups in the globulin. At pH 6.0, phaseolin is partly resistant to heat denaturation due to aggregation in an acidic environment. Isoelectrofocusing (IEF) and SDS polyacrylamide gel electrophoresis (SDS-PAGE) separated phaseolin into 10 protein fractions. The hypothesis is that phaseolin or globulin aggregation is due to the charge difference of fractions.     

大豆7S球蛋白亚基组成对品质性状的影响

以7S球蛋白亚基组成各异的稀有大豆种质为试材, 分析大豆7S球蛋白亚基组成对大豆营养品质性状的影响, 探索优质大豆品种选育的新思路。结果表明, 大豆7S球蛋白亚基组成对蛋白质、脂肪、蛋脂总量以及除缬氨酸和异亮氨酸外的15种氨基酸和5种脂肪酸含量都有一定的影响,对大豆籽粒内半胱氨酸含量的影响较大。另外, 各种亚基缺失型大豆材料的油酸、亚油酸和硬脂酸含量与该系统内正常型大豆材料的相应指标间的差异显著。