鸡Ii结合Rab5a和Rab7b分子的分析

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。     

Impact of grassland contract policy on soil organic carbon losses from alpine grassland on the Qinghai–Tibetan Plateau

Carbon storage in the soils on the Qinghai–Tibetan Plateau plays a very important role in the global carbon budget. In the 1990s, a policy of contracting collective grasslands to smaller units was implemented, resulting in a change from the traditional collective grassland management to two new management patterns: a multi‐household management pattern (MMP: grassland shared by several households without enclosures) and a single‐household management pattern (SMP: grassland enclosed and used by only one household). In 2016, 50 MMP and 54 SMP winter pastures on the Qinghai–Tibetan Plateau were sampled to assess the differences in soil organic carbon (SOC) between the two management patterns. Results showed that average SOC was significantly greater under MMP than under SMP, with an estimated 0.41 Mg C/ha/yr lost due to SMP following the new grassland contract. Based on the government's grassland policy, four grassland utilization scenarios were developed for both summer and winter pastures. We found that if the grassland were managed under SMP, likely C losses ranged between 0.31 × 10⁷ and 6.15 × 10⁷ Mg C/yr across the Qinghai–Tibetan Plateau relative to MMP, which more closely resembles pre‐1990s grassland management. Previous estimates of C losses have only considered land use change (with cover change) and ignored the impacts driven by land management pattern changes (without cover change). The new data suggest that C losses from the Qinghai–Tibetan Plateau are greater than previously estimated, and therefore that the grassland contract policy should be reviewed and SMP households should be encouraged to reunite into the MMP. These findings have potential implications for land management strategies not only on the Qinghai–Tibetan Plateau but also other grazing regions globally where such practices may exist.     

天敌昆虫低温贮藏研究进展

在农业生产中由于化学农药的长期不合理使用,导致"3R"问题持续存在和人类赖以生存的环境日益恶化,生物防治产品及技术在有害生物综合治理中的作用越发重要。天敌昆虫的生长发育及存活期往往很短,探索其储存技术是天敌昆虫规模化生产中的关键性环节。低温贮藏可以延长天敌昆虫的寿命,进而可以为生物防治提供充足、稳定的天敌产品。因此,天敌昆虫低温贮藏一直是生物防治领域研究的热点。本文围绕天敌昆虫低温贮藏这一环节,就低温诱导天敌昆虫滞育虫态、冷驯化对昆虫低温贮藏效果的影响、低温贮藏对天敌昆虫适合度(寿命、羽化率、性比、个体尺寸大小、飞行扩散能力等)的影响等方面进行综述,以期为天敌昆虫产品的规模化生产及应用技术提供参考。     

导入AGPase基因的转基因可育水稻及其经济性状的研究

以农杆菌介导法将淀粉合成关键酶AGPase(腺嘌呤二磷酸葡萄糖焦磷酸化酶）基因导入云南地方优质水稻合系35，38，39，41，42中，获得了大量的可育转基因植株。PCR扩增检测，外源基因已稳定整合进水稻基因组中。大量的农艺性状及淀粉含量研究表明，转基因水稻种子的千粒重比对照明显提高，但总淀粉含量没有显著变化。     

Studies on Tomato Seedling Quality Indices Under Simulated Shipping and Storage Conditions

Indices of the tomato seedling quality maintenance level after production before field planting were studied through simulated experiments, small--scale operation, indoor analyses and measurements, and field observation. The results showed that under simulated shipping and storage conditions, seedling quality change following different durations (days) of shipping and storage was correlated significantly or even very significantly with certain physiological and morphological indices. With various measured indices following different periods of shipping and storage treatment subjected to multinomial successive regressive correlation analysis, the principal factors influencing seedling quality maintenance level are identified to be chlorophyll content→dry short weight→ leaf freshness index in order of their importance. Significance analysis with multinomial fitted equation indicated that correlations between any one of above three factors and the growth index after field planting reached very significant difference level.     

分泌抗鸡Ig单抗杂交瘤细胞系的建立与单抗生物学特性的鉴定

从传染性法氏囊病病毒（ＩＢＤＶ）单克隆抗体（ＭｃＡｂ）杂交瘤细胞系１Ｄ１０、１Ｇ１中得到了３株能稳定分泌抗鸡免疫球蛋白（Ｉｇ）ＭｃＡｂ的杂交瘤细胞系１Ｂ７、１Ｄ７、３Ｇ６。其抗体类和亚类均属ＩｇＧ２ｂ，腹水的ＥＬＩＳＡ效价≥１∶２５６００，琼扩效价为１∶８～１∶１６。３株ＭｃＡｂ能与鸡血清中的Ｉｇ出现沉淀反应，琼扩在２４ｈ以内出现清晰的沉淀线，而不能和鸭、鸽、鹌鹑等禽类及异种动物血清出现沉淀线。纯化的鸡ＩｇＧ、ＩｇＭ经ＳＤＳ－ＰＡＧＥ后，分别用纯化并经辣根过氧化物酶（ＨＰＲ）标记的１Ｂ７、１Ｄ７、３Ｇ６单抗酶结合物进行免疫印迹试验证明，３株单抗识别的均为ＩｇＧ、ＩｇＭ分子的轻链     

甲苯侧链催化氯化产物分布的研究

研究了催化氯化的产物分布，试验了有关条件对产物分布的影响。结果表明：氯化程度是产物分布的主要影响因素；温度的提高，有利于苄叉二氯和苄川三氯的生成；铁离子、低温及水会导致环上的氯化；水引起水解的发生。     

Feline Ubiquitin Fusion Protein Genes

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.     

三分域模型在研究单胃动物内源氨基酸排泄中的应用

本文在分析三分域链状系统模型及单胃动物内源氨基酸排泄特点的基础上 ,讨论了如何将三分域链状模型用于单胃动物内源氨基酸排泄模式的确定。研究分三步进行 :1.测定动物血液体积 ;2.静脉连续灌注同位素标记的氨基酸并使之于体内达到平衡 ;3.检测消化道中同位素标记氨基酸的排泄率。文章指出 ,若能对所有必需氨基酸的内源排泄率均进行测定 ,则可得出内源氨基酸排泄模式 ,因而在实际测定饲料或日粮真可消化氨基酸含量时 ,只需测定某种氨基酸的内源排泄量即可知其它氨基酸的内源排泄状况     

RT-PCR及RFLP分析技术对鸡传染性支气管炎病毒的诊断和S1基因分型的研究

本研究使用分别扩增整个S1糖蛋白基因 (引物A)和S1糖蛋白基因N_端高变区Ⅰ (引物B)的 2对引物对 3个IBV标准株M41、Connecticut、Arkansas及 5个地方分离株 (C60 ,D41A ,D41B ,A112 1,A1171)进行RT_PCR扩增。用引物A时 ,有 5个IBV毒株扩增到目的片段 (172 0bp) ;用引物B时 ,所有 8个IBV毒株均得到与预计大小相符的目的产物 (2 2 8bp)。对 172 0bp的PCR产物用限制性内切酶HaeⅢ进行酶切 ,结果得出 3个不同的RFLP图谱 ,其中M41、Connecticut、D41B具有相同的HaeⅢ酶切图谱。Arkansas和D41A则分别具有互不相同的图谱 ;对 2 2 8bp的PCR产物进行DdeⅠ、RsaⅠ限制性内切酶消化 ,根据它们的RFLP图谱 ,8个IBV毒株可分为 5个基因型。综合 2对引物的PCR产物的RFLP分析结果 ,8个IBV毒株可分为 7个基因型 ,分型的结果与传统的血清学方法吻合。本研究建立的方法和技术具有快速、简单、特异、灵敏等优点 ,为现场流行毒株的定型(基因型 /血清型 )及其S1基因变异的跟踪研究以及更有效防制传染性支气管炎奠定了基础。