Ethylene has been implicated as a sex-determining hormone in cucumber. Its exogenous application increases femaleness,and gynoecious genotypes were reported to produce more ethylene. 1-aminocyclopropane-1-carboxylate ...Ethylene has been implicated as a sex-determining hormone in cucumber. Its exogenous application increases femaleness,and gynoecious genotypes were reported to produce more ethylene. 1-aminocyclopropane-1-carboxylate (ACC) oxidase(ACO) is the key enzyme in ethylene biosynthesis. In this study, a 1200 base pair (bp) candidate fragment was amplifiedfrom the cucumber genome with degenerated primers derived from the ACO amino acid consensus sequence amongdifferent plant species. The coding region and its upstream (1 155 bp) were obtained by vector-mediated inverse PCR. Thenovel gene was analyzed by bioinformatics tools. Four exons and three introns were identified in the coding sequence.The spliced length of mRNA was 933 nucleotides (nts) and it encoded 311 amino acids. Phylogenic analysis result of thenew gene (CsACO4, GenBank accession number AY450356) was in accordance with the evolution relationship of geneticsamong various plant species. Northern blotting showed that the gene expressed among female flowers of gynoecious andmonoecious genotypes, it could not express in other organs. This implied that the gene might be correlated with the femalebehavior positively. Further work is on the way to demonstrate the complexity of the relationship between the endogenousethylene and the sex determination.展开更多
In the present study, a full-length cDNA encoding 1-aminocyclopropane-1-carboxylic acid oxidase gene has been cloned from sugarcane （named GZ-ACO）. Two primers were designed for coding the ORF in the full-length cDN...In the present study, a full-length cDNA encoding 1-aminocyclopropane-1-carboxylic acid oxidase gene has been cloned from sugarcane （named GZ-ACO）. Two primers were designed for coding the ORF in the full-length cDNA of GZ-ACO gene from sugarcane. PCR amplification was performed with sugarcane DNA template, and a fragment of 1 104 bp （GZ34） was obtained. GZ34 was labeled with [α-32P] dCTP as the probe and used for hybridization after cloning and sequencing. Southern blotting analysis indicated that there were at least three other sequences, which weakly hybridized with the GZ34. Northern analysis showed that GZ34 was strongly induced by treatment with IAA, BA, ethephon, LiC1 and cold stress, respectively. As a contrast, the mRNA for ACO gene was at lower levels for both the light-grown and dark-grown plants without additional treatment. There were two transcripts in the dark-grown plants and three transcripts in the treatments with IAA, BA and cold stress, but there was only one transcript in ethephon treatment. It showed that GZ-ACO might be a gene connected with ethylene formation and take part in response to the induction of plant hormone and environmental stress.展开更多
Objective:To explore the influence of extract of Ginkgo biloba（EGB） on the proliferation, apoptosis of ACC-2 cell and Survivin gene expression in adenoid cystic carcinoma（ACC） of lacrimal gland.Methods:ACC-2 cell ...Objective:To explore the influence of extract of Ginkgo biloba（EGB） on the proliferation, apoptosis of ACC-2 cell and Survivin gene expression in adenoid cystic carcinoma（ACC） of lacrimal gland.Methods:ACC-2 cell in human with ACC of lacrimal gland was in vitro cultured. MTT method was used for cell proliferation detection.Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle.Survivin gene expression was analyzed by RT-PCR and Western blotting.Results:EGB had inhibitory effect on the proliferation of ACC-2 cell with significant dose-effect relationship,and there was statistical difference when compared with the control group（P【0.01）.The inhibitory concentration 50%(IC<sub>50</sub>) is 88 mg/L. The flow cytometer test indicated that CGB can gradually increase ACC-2 cell in G<sub>0</sub>-G<sub>1</sub> stage and decrease it in G<sub>2</sub>-M and S stage.With the increase of dose,the apoptosis rate of ACC-2 cell was obviously increased（P【0.05 or P【0.01）.EGB had certain inhibitor)’ effect on Survivin gene expression of ACC-2 cell,and Survivin gene expression was decreased with the increasing of the EGB concentration（P【0.01）.Conclusions:EGB can effectively inhibit Survivin gene expression of ACC-2 cell in human with ACC of lacrimal gland,induce the apoptosis of ACC-2 cell and inhibit tumor cell proliferation.展开更多
基金supported by the National Natural Science Foundation of China(39770521,39200079)
文摘Ethylene has been implicated as a sex-determining hormone in cucumber. Its exogenous application increases femaleness,and gynoecious genotypes were reported to produce more ethylene. 1-aminocyclopropane-1-carboxylate (ACC) oxidase(ACO) is the key enzyme in ethylene biosynthesis. In this study, a 1200 base pair (bp) candidate fragment was amplifiedfrom the cucumber genome with degenerated primers derived from the ACO amino acid consensus sequence amongdifferent plant species. The coding region and its upstream (1 155 bp) were obtained by vector-mediated inverse PCR. Thenovel gene was analyzed by bioinformatics tools. Four exons and three introns were identified in the coding sequence.The spliced length of mRNA was 933 nucleotides (nts) and it encoded 311 amino acids. Phylogenic analysis result of thenew gene (CsACO4, GenBank accession number AY450356) was in accordance with the evolution relationship of geneticsamong various plant species. Northern blotting showed that the gene expressed among female flowers of gynoecious andmonoecious genotypes, it could not express in other organs. This implied that the gene might be correlated with the femalebehavior positively. Further work is on the way to demonstrate the complexity of the relationship between the endogenousethylene and the sex determination.
基金the National Natural Science Foundation of China(39860039).
文摘In the present study, a full-length cDNA encoding 1-aminocyclopropane-1-carboxylic acid oxidase gene has been cloned from sugarcane （named GZ-ACO）. Two primers were designed for coding the ORF in the full-length cDNA of GZ-ACO gene from sugarcane. PCR amplification was performed with sugarcane DNA template, and a fragment of 1 104 bp （GZ34） was obtained. GZ34 was labeled with [α-32P] dCTP as the probe and used for hybridization after cloning and sequencing. Southern blotting analysis indicated that there were at least three other sequences, which weakly hybridized with the GZ34. Northern analysis showed that GZ34 was strongly induced by treatment with IAA, BA, ethephon, LiC1 and cold stress, respectively. As a contrast, the mRNA for ACO gene was at lower levels for both the light-grown and dark-grown plants without additional treatment. There were two transcripts in the dark-grown plants and three transcripts in the treatments with IAA, BA and cold stress, but there was only one transcript in ethephon treatment. It showed that GZ-ACO might be a gene connected with ethylene formation and take part in response to the induction of plant hormone and environmental stress.
文摘Objective:To explore the influence of extract of Ginkgo biloba（EGB） on the proliferation, apoptosis of ACC-2 cell and Survivin gene expression in adenoid cystic carcinoma（ACC） of lacrimal gland.Methods:ACC-2 cell in human with ACC of lacrimal gland was in vitro cultured. MTT method was used for cell proliferation detection.Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle.Survivin gene expression was analyzed by RT-PCR and Western blotting.Results:EGB had inhibitory effect on the proliferation of ACC-2 cell with significant dose-effect relationship,and there was statistical difference when compared with the control group（P【0.01）.The inhibitory concentration 50%(IC<sub>50</sub>) is 88 mg/L. The flow cytometer test indicated that CGB can gradually increase ACC-2 cell in G<sub>0</sub>-G<sub>1</sub> stage and decrease it in G<sub>2</sub>-M and S stage.With the increase of dose,the apoptosis rate of ACC-2 cell was obviously increased（P【0.05 or P【0.01）.EGB had certain inhibitor)’ effect on Survivin gene expression of ACC-2 cell,and Survivin gene expression was decreased with the increasing of the EGB concentration（P【0.01）.Conclusions:EGB can effectively inhibit Survivin gene expression of ACC-2 cell in human with ACC of lacrimal gland,induce the apoptosis of ACC-2 cell and inhibit tumor cell proliferation.
