山杏苹果酸酶基因的克隆与生物信息学分析

三羧酸转运体系(柠檬酸,苹果酸和丙酮酸)可为脂肪酸生物合成提供原料乙酰辅酶A与还原力NADPH;而苹果酸酶可催化苹果酸发生氧化脱羧而产生NADPH,是调控脂肪酸代谢的重要酶。山杏种子脂肪酸含量丰富、产油率高,是研究植物油脂代谢的良好材料。本研究以山杏为试材,采用RACE克隆和电子拼接相结合的方法,首次从山杏种子中克隆得到苹果酸酶基因(命名为SaME,GenBank上登记号为JX262381),该基因的cDNA编码区全长1923bp、编码640个氨基酸,预测其编码蛋白分子量为70.15kD、等电点为6.38;同时,运用生物信息学方法对SaME基因编码蛋白的理化特性、结构域和亚细胞定位等方面进行预测分析,结果显示,该蛋白定位在细胞的质体膜上,具有苹果酸酶活性,属于NADP-ME超家族。本文对山杏SaME基因的克隆与分析,为进一步探究苹果酸酶对油脂代谢的调控机制奠定了基础。     

Microbe-derived non-necrotic glycoside hydrolase family 12 proteins act as immunogenic signatures triggering plant defenses

Plant pattern recognition receptors(PRRs) are sentinels at the cell surface sensing microbial invasion and activating innate immune responses. During infection, certain microbial apoplastic effectors can be recognized by plant PRRs, culminating in immune responses accompanied by cell death. However, the intricated relationships between the activation of immune responses and cell death are unclear.Here, we studied the glycoside hydrolase family12(GH12) protein, Ps109281, secreted by Phytophthora ...     

模拟喀斯特不同土壤生境下黑麦草对水分胁迫的生长和光合生理响应

以多年生草本黑麦草为对象,模拟喀斯特生境土壤特征设置浅而宽(Shallow and wide, SW:30×30×5 cm~3)和深而窄(Deep and narrow, DN:10×10×45 cm~3)两种土壤容器,以正常降水量为对照供水(W100%),设减水50%(W50%)和减水70%(W30%)共3种水分处理进行盆栽实验,探究了两种不同土壤生境中土壤水分变化对黑麦草生长及光合生理的影响,以进一步理解喀斯特地区植物的适应对策。结果显示:(1)SW生境对水分变化敏感,随供水减少土壤含水量显著下降。轻度减水下植物叶面积增大,光合速率提高,地上部分生长得到促进,但水分严重减少对其生长和光合生理有抑制作用,但地上质量分数和水分利用效率却显著升高;(2)DN生境保水能力较好,随供水减少土壤水分含量下降较为平缓。叶片相对含水量、气孔导度和比叶面积在各水分处理之间差异不显著,但严重减水条件下总生物量、地上质量分数和水分利用效率均有回升。研究表明:浅而宽生境中植物倾向于通过提高地上部分的生长,保持较高的光合速率,并向地上部分分配较多生物量来应对水分胁迫;而深而窄生境中植物会充分利用土壤空间条件,促进根系的生长来提高对水分的吸收,从而维持植物的生长。     

基于DTTD-MCR-PLUS模型的三生空间格局优化——以长沙市为例

优化三生空间格局有助于实现区域国土空间可持续高质量发展。研究提出利用动态交通时间数据优化最小阻力成本模型指标体系,对生态累计最小阻力与生活最小累计阻力差值进行聚类分析,利用突变点对各情景下的三生空间数量进行优化,从生活扩张、生态优先及基于资源与环境承载力视角下的粮食安全3个方面,提出了一种基于DTTD-MCR-PLUS的三生空间格局优化方法。研究发现：1) 基于动态数据优化的长沙市生态功能优化分区结果显示生态空间保护核心区面积为4111.41 km2,生态空间保护边缘区面积为2285.29 km2,生产空间开发重点区面积为2144.79 km2,生产空间开发边缘区面积为1928.59 km2,生活空间扩张集中区面积为1235.55 km2; 2) 耦合DTTD-MCR-PLUS模型模拟的多情景结果表明：生活优先情景下,生活空间面积增幅高达43.57%,主要分布在望城区南部,长沙县西部和雨花区东部;生态优先情景下,生态空间转出速度最低,与生活优先情景相比下降了3.11%;粮食安全情景下,生产空间侵占生态空间速度加快,增幅高达58.79%;3) 协调基本农田、生态保护红线、以及自然保护区下的2030 年长沙市三生空间格局优化布局方案结果表明：生产空间、生活空间和生态空间比例分别为37.63%、7.67%和54.70%。     

水杨酸对铜胁迫下菊芋的生理调控及根系FTIR分析

为阐明水杨酸(salicylic acid,SA)对菊芋吸收铜的影响机理,以耐铜性差异较大的徐州菊芋(Helianthus tubeuosus)和潍坊菊芋作为试验材料,通过盆栽试验,探究外源SA对铜胁迫下菊芋的光合作用、叶片抗氧化系统及必需矿质营养元素吸收的影响,并对其耐铜机理进行探究。结果表明,300mg/kg铜胁迫抑制了两地菊芋光合电子传递效率和PSⅡ活性,对抗氧化系统产生损伤并造成膜脂过氧化,显著增加了根、茎、叶中铜含量,同时削弱了菊芋对钾、钙、镁和锌的吸收能力。喷施1 mmol/L外源SA可通过调控根系中各官能团的比例,加强其对Cu^(2+)的固定作用,促进矿质营养元素的吸收及渗透调节物质的积累,提高菊芋抗氧化和光合作用能力,有效缓解铜对菊芋生长的抑制作用。本研究初步揭示了SA对铜胁迫下菊芋的生理调控,可为种植菊芋修复土壤铜污染提供科学参考和依据。     

EcoRⅡ甲基化酶基因在小鼠NIH3T3细胞中的表达

CG外甲基化在哺乳动物细胞中的生物学意义仍不清楚。建立了特定位点CCWGG（W＝A／T）甲基化的哺乳动物细胞株。来自大肠杆菌的EcoRⅡ甲基化酶（催化CCWGG→CmCWGG）基因经定点诱变后重组到真核表达载体pCI－neo上,采用脂质体介导的方法将其转染小鼠NIH3T3细胞,经G418筛选获得有效表达的细胞株,酶切证实该细胞株基因组DNA上的大部分EcoRⅡ识别位点已被甲基化,PCR鉴定EcoRⅡ甲基化酶基因已稳定整合到细胞染色体上。     

A simple microsphere-based mold to rapidly fabricate microwell arrays for multisize 3D tumor culture

Three-dimensional (3D) tumor has been considered as the best in vitro model for cancer research. In recent years, various methods have been developed to controllable prepare multisize 3D tumors. Nonetheless, reported technologies are still problematic and difficult to produce 3D tumors with highly uniform size and cell content. Here, a novel and simple microsphere-based mold approach is proposed to rapidly fabricate spherical microwell arrays for multisize 3D tumors formation, culture, and recovery. Larger amounts of HepG2 3D tumors with excellent quality and uniformity can be efficiently generated using this method. In addition, the tumor size can also be simply controlled by adjusting the diameter of the microwell arrays. All experimental results indicated that the proposed method offers a promising platform to generate and recover highly controlled multisize 3D tumors for various cell-based biomedical research.     

Identification of key biomarkers for thyroid cancer by integrative gene expression profiles

Thyroid cancer is a frequently diagnosed malignancy and the incidence has been increased rapidly in recent years. Despite the favorable prognosis of most thyroid cancer patients, advanced patients with metastasis and recurrence still have poor prognosis. Therefore, the molecular mechanisms of progression and targeted biomarkers were investigated for developing effective targets for treating thyroid cancer. Eight chip datasets from the gene expression omnibus database were selected and the inSilicoDb and inSilicoMerging R/Bioconductor packages were used to integrate and normalize them across platforms. After merging the eight gene expression omnibus datasets, we obtained one dataset that contained the expression profiles of 319 samples (188 tumor samples plus 131 normal thyroid tissue samples). After screening, we identified 594 significantly differentially expressed genes (277 up-regulated genes plus 317 down-regulated genes) between the tumor and normal tissue samples. The differentially expressed genes exhibited enrichment in multiple signaling pathways, such as p53 signaling. By building a protein–protein interaction network and module analysis, we confirmed seven hub genes, and they were all differentially expressed at all the clinical stages of thyroid cancer. A diagnostic seven-gene signature was established using a logistic regression model with the area under the receiver operating characteristic curve (AUC) of 0.967. Seven robust candidate biomarkers predictive of thyroid cancer were identified, and the obtained seven-gene signature may serve as a useful marker for thyroid cancer diagnosis and prognosis.     

Guanine nucleotide-binding protein subunit beta-2-like 1, a new Annexin A7 interacting protein

We report for the first time that Guanine nucleotide-binding protein subunit beta-2-like 1 (RACK1) formed a complex with Annexin A7. Hca-F and Hca-P are a pair of syngeneic mouse hepatocarcinoma cell lines established and maintained in our laboratory. Our previous study showed that both Annexin A7 and RACK1 were expressed higher in Hca-F (lymph node metastasis >70%) than Hca-P (lymph node metastasis <30%). Suppression of Annexin A7 expression in Hca-F cells induced decreased migration and invasion ability. In this study, knockdown of RACK1 by RNA interference (RNAi) had the same impact on metastasis potential of Hca-F cells as Annexin A7 down-regulation. Furthermore, by co-immunoprecipitation and double immunofluorescence confocal imaging, we found that RACK1 was in complex with Annexin A7 in control cells, but not in the RACK1-down-regulated cells, indicating the abolishment of RACK1-Annexin A7 interaction in Hca-F cells by RACK1 RNAi. Taken together, these results suggest that RACK1-Annexin A7 interaction may be one of the means by which RACK1 and Annexin A7 influence the metastasis potential of mouse hepatocarcinoma cells in vitro.