松墨天牛dynamin-1-like protein基因的鉴定及表达分析

为了探讨GTP酶超基因家族中dynamin-1-like protein基因在昆虫中的表达特性,以松墨天牛为研究对象,从已构建的cDNA文库中筛选到松墨天牛dynamin-1-like protein基因,命名为Ma DLP1(Gen Bank:KU245763)。该序列长为2 133 bp,编码710个氨基酸。由此预测的蛋白质二级结构主要由α螺旋与无规则卷曲组成,其次是β片层与β转角;Ma DLP1基因编码蛋白质,定位于细胞核。通过DNAMAN软件比对发现Ma DLP1与赤拟谷盗的DLP1同源性最高,为82%,且存在3个保守酶域;用Clustal X和MEGA4.0构建系统发育树,显示松墨天牛与赤拟谷盗处在同一分支。RT-qPCR分析结果显示,Ma DLP1在各虫态不间断表达,幼虫期在5龄幼虫中表达量最高,化蛹期间表达量先上升后下降,羽化期间表达量表现为先上升后下降,并在初羽化的成虫中表达量达到最大值;Ma DLP1在幼虫和成虫头部表达量较高,且在幼虫脂肪体中表达最高;成虫的足、翅、触角和卵巢中也都有表达。说明Ma DLP1的表达与松墨天牛的完全变态发育相关。

与肌少症发病相关的线粒体自噬靶点基因筛选及其在骨骼肌组织中表达观察

目的基于GEO数据库数据筛选与肌少症发病相关的线粒体自噬靶点基因,并观察其在肌少症患者骨骼肌组织中的表达变化。方法从GEO数据库检索肌少症的基因图谱数据,筛选肌少症发病的差异表达基因。从GeneCard数据库中检索并收集线粒体自噬相关基因。使用“VennDiagram”包将肌少症发病的差异表达基因与线粒体自噬相关基因取交集,得到与肌少症发病相关的线粒体自噬差异表达基因。运用基因本体论(GO)和京都基因与基因百科全书(KEGG)通路富集分析与肌少症发病相关的线粒体自噬差异表达基因的生物学功能,通过Cytoscape软件筛选与肌少症发病相关的线粒体自噬靶点基因,观察GSE136344基因表达图谱中肌少症、健康对照者骨骼肌与肌少症发病相关的线粒体自噬靶点基因表达情况。结果得到与肌少症发病相关的线粒体自噬差异表达基因99个。与肌少症发病相关的线粒体自噬差异表达基因主要涉及神经变性途径-多种疾病信号通路、帕金森疾病信号通路、朊毒体病信号通路等;主要调控能量代谢、细胞呼吸、氧化磷酸化调节等生物学过程,主要定位于线粒体内膜、线粒体内部的大分子蛋白质复合物等,参与调节跨膜转运活性等分子功能。与肌少症发病相关的线粒体自噬靶点基因有线粒体内膜蛋白基因(IMMT)、动态蛋白1样蛋白基因(DNM1L)及ATP合酶F1亚基α基因(ATP5A1)等;与正常骨骼肌组织相比,肌少症患者骨骼肌组织中IMMT、DNM1L表达低(P均<0.05)。结论与肌少症发病相关的线粒体自噬靶点基因为IMMT、DNM1L。与肌少症发病相关的线粒体自噬靶点基因可通过影响神经变性途径-多种疾病信号通路、帕金森疾病信号通路及朊毒体病信号通路等,参与调控能量代谢、细胞呼吸、氧化磷酸化调节等生物学过程,参与肌少症的发病。肌少症患者骨骼肌组织中IMMT、DNM1L低表达。

Genome-wide identification of abscisic acid(ABA) receptor pyrabactin resistance 1-like protein(PYL) family members and expression analysis of PYL genes in response to different concentrations of ABA stress in Glycyrrhiza uralensis

As abscisic acid(ABA)receptor,the pyrabactin resistance 1-like(PYR/PYL)protein(named PYL for simplicity)plays an important part to unveil the signal transduction of ABA and its regulatory mechanisms.Glycyrrhiza uralensis,a drought-tolerant medicinal plant,is a good model for the mechanism analysis of ABA response and active compound biosynthesis.However,knowledge about PYL family in G.uralensis remains largely unknown.Here,10 PYLs were identified in G.uralensis genome.Characterization analysis indicated that PYLs in G.uralensis(Gu PYLs)are relatively conserved.Phylogenetic analysis showed that Gu PYL1-3 belongs to subfamily I,Gu PYL4-6 and Gu PYL10 belong to subfamily II and Gu PYL7-9 belongs to subfamily III.In addition,transcriptome data presented various expression levels of Gu PYLs under different exogenous ABA stresses.The expression pattern of Gu PYLs was verified by Quantitative real-time polymerase chain reaction(q RT-PCR).The study proved that Gu PYL4,Gu PYL5,Gu PYL8 and Gu PYL9 genes are significantly up-regulated by ABA stress and the response process is dynamic.This study paves the way for elucidating the regulation mechanism of ABA signal to secondary metabolites and improving the cultivation and quality of G.uralensis using agricultural strategies.

Interaction of the major inflammatory bowel disease susceptibility alleles in Crohn’s disease patients

AIM:To investigate the interaction of interleukin-23 receptor(IL23R)(rs1004819 and rs2201841),autophagy-related 16-like 1(ATG16L1)(rs2241880), caspase recruitment domain-containing protein 15 (CARD15)genes,and IBD5 locus in Crohn's disease(CD) patients. METHODS:A total of 315 unrelated subjects with CD and 314 healthy controls were genotyped.Interactions and specific genotype combinations of a total of eight variants were tested.The variants of IBD5locus(IGR2198a_1 rs11739135 and IGR2096a_1 rs12521868),CARD15(R702W rs2066845 and L1007fs rs2066847),ATG16L1(rs2241880)and IL23R (rs1004819,rs2201841)genes were genotyped by PCR-RFLP,the G908R(rs2066844)in CARD15 was determined by direct sequencing. RESULTS:The association of ATG16L1 T300A with CD was confirmed[P=0.004,odds ratio(OR)=1.69, 95%CI:1.19-2.41],and both IL23R variants were found to represent significant risk for the disease(P= 0.008,OR=2.05,95%CI:1.20-3.50 for rs1004819 AA;P<0.001,OR=2.97,95%CI:1.65-5.33 for rs2201841 CC).Logistic regression analysis of pairwise interaction of the inflammatory bowel disease (IBD)loci indicated that IL23R,ATG16L1,CARD15 and IBD5(IGR2198a_1)contribute independently to disease risk.We also analysed the specific combina- tions by pair of individual ATG16L1,IL23R rs1004819, rs2201841,IGR2198a_1,IGR2096a_1 and CARD15 genotypes for disease risk influence.In almost all cases,the combined risk of susceptibility pairs was higher in patients carrying two different risk-associated gene variants together than individuals with just one polymorphism.The highest OR was found for IL23R rs2201841 homozygous genotype with combination of positive CARD15 status(P<0.001,OR=9.15,95% CI:2.05-40.74). CONCLUSION:The present study suggests a cumulative effect of individual IBD susceptibility loci.