独行菜eEF-1a基因片段分离克隆及RT-PCR分析

文章根据已知植物eEF-1a(编码Eukaryotic elongation factor1-alpha)基因的保守序列设计引物,采用RT-PCR的方法扩增独行菜(Lepidium apetalumWilld.)eEF-1a基因片段。结果获得一大小为570bp的基因片段,序列比对表明,该基因片段与拟南芥eEF-1a基因核苷酸序列的同源性达到95%,推测该其应该是独行菜eEF-1a基因片段。采用RT-PCR方法,对独行菜三个生长阶段材料及对应阶段冷诱导后地材料中的eEF-1a基因表达作了分析,表明eEF-1a在独行菜组织中表达稳定,可以作为实时荧光定量PCR研究独行菜基因表达分析中的看家基因。

Effects of eEF1A1 expression on cancer cell load and proliferation activity in lung cancer lesions

Objective: To investigate the effects of eEF1A1 expression on cancer cell load and proliferation activity in lung cancer lesions. Methods: A total of 200 patients with non-small cell lung cancer who received radical operation for lung cancer in our hospital were enrolled in lung cancer group, and 78 patients with pulmonary bullae who accepted emergency surgical treatment in our hospital during the same period were enrolled in pulmonary bullae group. Differences in eEF1A1 and proliferation gene expression in lesion tissue were compared between the two groups, and the contents of serum tumor markers and angiogenesis indexes were detected. Pearson test was used to evaluate the correlation of eEF1A1 expression with tumor markers, angiogenesis indexes and proliferation genes in the lesions of patients with non-small cell lung cancer. Results: eEF1A1 expression in lesion tissue of lung cancer group was higher than that of pulmonary bullae group;serum CYFRA21-1, ProGRP, CEA, SCC Ag, HIF-1 and VEGF levels of lung cancer group were higher than those of pulmonary bullae group and positively correlated with the eEF1A1 expression;LRRC3B, TUSC3 and VPS33B mRNA expression in lesion tissue of lung cancer group were lower than those of pulmonary bullae group and negatively correlated with the eEF1A1 expression whereas MACC1 and RACK1 mRNA expression were higher than those of pulmonary bullae group and positively correlated with the eEF1A1 expression. Conclusion: eEF1A1 is highly expressed in non-small cell lung cancer tissues, and the specific expression can objectively reflect the tumor load and the proliferation activity of tumor cells.

LMM5.1 and LMM5.4,two eukaryotic translation elongation factor 1Alike gene family members,negatively affect cell death and disease resistance in rice

Lesion mimic mutant（LMM） genes, stimulating lesion formation in the absence of pathogens, play significant roles in immune response. In this study, we characterized a rice lesion mimic mutant, lmm5,which displayed light-dependent spontaneous lesions. Additionally, lmm5 plants exhibited enhanced resistance to all of the tested races of Magnaporthe oryzae and Xanthomonas oryzae pv. oryzae（Xoo） by increasing the expression of defense-related genes and the accumulation of hydrogen peroxide. Genetic analysis showed that the lesion mimic phenotype of lmm5 was controlled by two genes, lmm5.1 and lmm5.4, which were isolated with a map-based cloning strategy. Remarkably, LMM5.1 and LMM5.4 share a 97.4% amino acid sequence identity, and they each encode a eukaryotic translation elongation factor 1A（e EF1A）-like protein. Besides, LMM5.1 and LMM5.4 were expressed in a tissue-specific and an indicaspecific manner, respectively. In addition, high-throughput m RNA sequencing analysis confirmed that the basal immunity was constitutively activated in the lmm5 mutant. Taken together, these results suggest that the homologous e EF1A-like genes, LMM5.1 and LMM5.4, negatively affect cell death and disease resistance in rice.

mRNA差异显示技术分离羊草基因体外培养过程中特定基因及特性分析

羊草种质资源中储存着潜在的优良基因,本研究从分子生物学角度,对来自羊草基因型W4不同幼穗的愈伤组织中差异表达的基因进行了研究。采用DDRT-PCR技术对其差异表达的基因进行了分离,通过银染技术显示差异片段。将得到的差异片段进行回收、克隆测序,得到2个差异片段序列,经过序列分析表明,其中1个片段是与水稻翻译延伸因子eEF-1基因高度同源;另一差异片段与水稻谷胱甘肽转移酶GST基因高度同源。