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| 1 | Cloning and characterization of a new actin gene from Oryza sativa L.显示文摘Using Rho family member osRACD as bait,a new member of actin gene family-Act was isolated from Oryza sativa by yeast two-hybrid system.The full-length cDNA was cloned with5′RACE technology,which contains an open reading frame of1134bp with a predicted protein of377amino acids.Sequence alignment revealed96%to81.8%identities with some known actin proteins in plants.The method of bioinformatics was used to analyze the protein modification sites,structure and evolution of the gene.Southern blot analysis showed that Act is a single-copy gene in the genome.The result of RT-PCR showed it is ubiquitously expressed in root,shoot,callus and panicle in a temporal fashion.The relationship between Rho family and actin family in evolution and function was also studied. | LIANGWeihong TANGChaorong WUNaihu | 2004 | Progress in Natural Science:Materials International2004，14，10： | 18 |
| 2 | Fertility analysis of the Arabidopsis transformed with antisense rice osRACD gene显示文摘The full length osRACD cDNA sequence was subcloned into the pBI121 plasmid in the antisense orientation under the control of the CaMV35S promoter to construct the expression vector pBID, and the constructs were introduced into Arabidopsis plants by using the vacuum infiltration method. The siliques of the transformants stopped growing after anthesis, and they turned yellow or died later; and the siliques from the control plants transformed by the pBI continued growing after anthesis and matured normally. In vitro pollen germination demonstrated that the growth and elongation process of the pollens of the transgenic plants was inhibited, the pollen tubes were shorter and slightly fatter than the tubes of the control plants, which grew normally with long cyclindrical tubes. The above results suggest the function of osRACD gene involved in regulation of the growth and elongation process of pollen tube, its encoding protein may be one of the important factors in regulation of fertility transition of the photoperiod-sensitive genic male-sterile rice Nongken 58S. | YEJianrong HUANGMeijuan WUNaihu | 2003 | Progress in Natural Science:Materials International2003，13，6： | 8 |
| 3 | Molecular cloning of C4-specific Ppc gene of sorghum and its high level expression in transgenic rice显示文摘In order to improve the carbon-assimilation ability of C3 plants, we isolated a C4-specific photosynthetic enzyme gene, Ppc (encode phosphoenolpyruvate carboxylase, PEPCase) from the genome of the C4 plant, sorghum, and transformed rice with it. As shown by sequence analysis, the gene is composed of 10 exons and 9 introns, and the full-length transcript is 5989 bp long. A recombinant expres-sion vector, p1301PEPC, was constructed by inserting the gene into a plasmid vector, pCAMBIA1301, which was then transformed into two japonica rice varieties, Nongken 58 and Zhonghua 10, using an Agrobacterium-mediated transforma-tion system. PCR analysis, activity measurement of PEPCase, and protein-, RNA- and DNA-based hybridization all con-firmed the successful integration of the C4-specific Ppc gene into the nuclear genome of rice and its high level expression. Physiological studies revealed the photosynthetic features characterizing C4 plants such as marked lowering of CO2 compensation point and photorespiration rate, and im-proved carboxylation efficiency. This study provides useful experimental materials and opens up new avenues for fur-ther studies on improving photosynthetic efficiency of elite varieties of rice. | ZHANGFang CHIWei WANGQiang ZHANGQide WUNaihu | 2003 | Chinese Science Bulletin2003，48，17： | 4 |
| 4 | Cloning and expression of DnaJ homolog in carrot somatic embryo显示文摘As the co-chaperone of DnaK/Hsp70 protein, DnaJ/Hsp40 protein influences the synthesis and assembly of the protein complex by regulating ATPase activity of DnaK/Hsp70 protein. By employing the modified method of cDNA representational difference analysis, a homologous fragment of DnaJ was isolated from the deregulated carrot somatic embryos, and it was further used as the probe to screen the cDNA library of carrot somatic embryo deregulated for 12 h. As the result, DcJ1 gene, the homologous gene of DnaJ, was isolated from carrot. Sequence analysis showed that its coding region is 1257 bp, which codes 418 amino acids and comprises 3 highly-conserved characteristic domains. Southern blot analysis suggested that the DcJ1 gene seems to be a single copy in the genome, while Northern blot result indicated that DcJ1 expresses only in roots and its degree of expression changes obviously with the regulation-deregulation process. These results suggest that DcJ1 is correlated with the early development of carrot somatic embryo radicle. | YANGZhipan ZHANGLei DIAOFengqiu HUANGMeijuan WUNaihu | 2003 | Progress in Natural Science:Materials International2003，13，5： | 0 |
| 5 | Cloning and characterization of DcLEA1,a new member of carrot LEA gene family显示文摘Using a modified cDNA representational difference analysis(RDA)method,a LEA gene fragment was isolated from the regulated carrot somatic embryo,which was used as the probe to screen the cDNA library of the regulated carrot somatic embryo and the genomic library constructed by the method of altering osmotic pressure.Sequence analysis showed that it is homologous to LEA gene family and designated as DcLEA1(GenBank number:AF308739),a new member of the carrot LEA gene family.Its transcription re-gion contains5′UTR,two exons,one intron and3′UTR region;its coding region is480bp long,coding for159amino acids and one stop codon.Northern hybridization indicated that DcLEA1gene was not expressed in the adult carrot but expressed at high levels in the regulated carrot somatic embryo.In carrot somatic embryo which had been deregulated for12hours,the expression levels dropped rapid-ly;with the prolongation of deregulation,the radicle of carrot somatic embryo began to stretch,and the expression level of DcLEA1gene increased.This phenomenon is similar to the expression pattern of LEA gene in the course of dormancy and germination of the seed;thus suggesting that the sucrose regulation-deregulation system of the carrot somatic embryo can be used to mimic plant seed dormancy and ger-mination and can also be used to study the molecular mechanisms of these two biological processes. | LIUYiming DIAOFengqiu ZHANGLei HUANGMeijuan WUNaihu | 2005 | Progress in Natural Science:Materials International2005，15，3： | 0 |
| 6 | Biochemical characterization of the protein encoded by rice osRACD gene显示文摘A recombinant plasmid pET-racd was first constructed by cloning osRACD, the development-regulating gene that controls photoperiod fertility transformation in the photoperiod sensitive genic male sterile rice Nongken 58S, into a prokaryote expression vector pET28a( + ). It was then transformed into E. coli BL-21. Cutting with thrombin of the fusion protein extracted from transforrnants and PAGE separation yielded pure osRACD protein, which was further concentrated using ultrafiltration and renatured using glutathione oxidation/reduction refolding system for later functional study. As demonstrated by in vitro functional assay, the osRACD protein expressed in E. coli B-21 shows remarkable activity in binding GTP specifically and hydrolyzing it. | XUYunzhe MIZhiyong QUHong YOURuilin WUNaihu | 2004 | Progress in Natural Science:Materials International2004，14，4： | 0 |
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