Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the...Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal Rhodophyta. Here, the effects of salinity and light intensity on the quantum yield of photosystem II in Py. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under different stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The cDNA was synthesized and the expression levels of seven potential internal control genes were evaluated using qRT-PCR method. Then, we used geNorm, a common statistical algorithm, to analyze the qRT-PCR data of seven reference genes. Potential genes that may constantly be expressed under different conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while tubulin, glyceraldehyde- 3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the geNorm screening. Based on the results of the software analysis and absolute quantification, we proposed that elongation factor 3 and 18S ribosomal RNA could be used as internal control genes when the Py. haitanensis blades were subjected to salinity stress, and that a-tubulin and 18S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis.展开更多
Nanosecond laser pulse photolysis was used to investigate the excited-state dynamics of the photonuclease vitamin-K3 (VK3). A spectral band peaked at 285 nm was newly observed and attributed to the triplet-excited sta...Nanosecond laser pulse photolysis was used to investigate the excited-state dynamics of the photonuclease vitamin-K3 (VK3). A spectral band peaked at 285 nm was newly observed and attributed to the triplet-excited state of VK3. In studying the photochemistry between the photonu-clease VK3 and guanine nucleotide (dGMP), we obtained spectroscopic evidence for the photo-initiated formation of both VK3 radical anion and dGMP radical cation, and thus proved the charge transfer reaction between the trip-let-excited VK3 and the ground-state dGMP.展开更多
In this paper, we report a highly sensitive chemiluminescence(CL) sensor for Hg2+ ions based on thymine-Hg2+-thymine(T-Hg2+-T) coordination chemistry. We designed a thymine rich oligonucleotide as a capture probe and ...In this paper, we report a highly sensitive chemiluminescence(CL) sensor for Hg2+ ions based on thymine-Hg2+-thymine(T-Hg2+-T) coordination chemistry. We designed a thymine rich oligonucleotide as a capture probe and a signal probe that includes two functional domains: a horseradish peroxidase-mimicking DNAzyme domain for the generation of CL, and a recognition domain. Graphene oxide(GO) was introduced to adsorb the signal probe via π-π interaction, which brought the DNAzyme domain and GO into close proximity and quenches CL. In the presence of Hg2+ ions, the coordination of Hg2+ with the capture probe yielded a hairpin complex, triggers cascaded strand displacement reactions and Exonuclease III-assisted signal amplifications. As a result, accumulated amounts of DNAzyme were generated and released from GO, leading to an enhanced CL signal. This strategy combines enzyme-based signal amplification and GO as a background reducer, leads to a limit of detection(LOD) of 2 nmol/L. This simple detection system provides a label-free yet sensitive approach for detection of Hg2+ ions.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.41476140,41306151,41676157,41506172)the Strategic Leading Science and Technology Projects of Chinese Academy of Sciences(No.XDA11020404)+1 种基金the China Postdoctoral Science Foundation(No.2015M582153)the Science and Technology Plan of Jiangsu Province(No.BE2016330)
文摘Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal Rhodophyta. Here, the effects of salinity and light intensity on the quantum yield of photosystem II in Py. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under different stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The cDNA was synthesized and the expression levels of seven potential internal control genes were evaluated using qRT-PCR method. Then, we used geNorm, a common statistical algorithm, to analyze the qRT-PCR data of seven reference genes. Potential genes that may constantly be expressed under different conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while tubulin, glyceraldehyde- 3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the geNorm screening. Based on the results of the software analysis and absolute quantification, we proposed that elongation factor 3 and 18S ribosomal RNA could be used as internal control genes when the Py. haitanensis blades were subjected to salinity stress, and that a-tubulin and 18S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis.
文摘Nanosecond laser pulse photolysis was used to investigate the excited-state dynamics of the photonuclease vitamin-K3 (VK3). A spectral band peaked at 285 nm was newly observed and attributed to the triplet-excited state of VK3. In studying the photochemistry between the photonu-clease VK3 and guanine nucleotide (dGMP), we obtained spectroscopic evidence for the photo-initiated formation of both VK3 radical anion and dGMP radical cation, and thus proved the charge transfer reaction between the trip-let-excited VK3 and the ground-state dGMP.
基金supported by the National Natural Science Foundation of China(21222508,21375073)the Shanghai Municipal Commission for Science and Technology(13QH1402300)+1 种基金the State Ethnic Affairs Commission(10ZY02)the 111 Project of Minzu University(B08044)
文摘In this paper, we report a highly sensitive chemiluminescence(CL) sensor for Hg2+ ions based on thymine-Hg2+-thymine(T-Hg2+-T) coordination chemistry. We designed a thymine rich oligonucleotide as a capture probe and a signal probe that includes two functional domains: a horseradish peroxidase-mimicking DNAzyme domain for the generation of CL, and a recognition domain. Graphene oxide(GO) was introduced to adsorb the signal probe via π-π interaction, which brought the DNAzyme domain and GO into close proximity and quenches CL. In the presence of Hg2+ ions, the coordination of Hg2+ with the capture probe yielded a hairpin complex, triggers cascaded strand displacement reactions and Exonuclease III-assisted signal amplifications. As a result, accumulated amounts of DNAzyme were generated and released from GO, leading to an enhanced CL signal. This strategy combines enzyme-based signal amplification and GO as a background reducer, leads to a limit of detection(LOD) of 2 nmol/L. This simple detection system provides a label-free yet sensitive approach for detection of Hg2+ ions.
