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Expression of the mouse metallothionein mutant ββ-cDNA in the lettuces (Lactuca sativa L.)

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Chinese Science Bulletin 2002年第7期47卷 558-562页

作者： Zuo Xiaofeng Zhang Yukun Wu Beibei Chang Xing Ru Binggen state key laboratory of protein engineering and plant genetic engineering Peking University100871BeijingChina

Mouse metallothionein (MT) domain mutant ββ-cDNA gene has been inserted into the plant expression vector pGPTVd35S which has herbicide (PPT)-resistance gene bar as a selective marker. The chimeric gene was introduced into the lettuce (Lactuca sativa L. cv Salinas 88) by Agrobacterium-mediated transformation. PCR and Southern blot analysis of some putative transformants indicated that the introduced ββ-cDNA was integrated into lettuce genome and inheritted by sexual reproduction. The expression of ββ gene in lettuce plants was demonstrated by Northern and Western blot analysis. Meanwhile, the zinc content of the transformed lettuce plants is up to 400 μg/g dry weight, remarkably higher than the control plants.

关键词： metallothionein domain mutant pp gene zinc content lettuce.

Molecular cloning, sequence analysis and expression studies of a novel GAmyb homologous gene, hvmyb

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Science China(Life Sciences) 1998年第5期41卷 525-529页

作者：凌世昀祁放朱玉贤 1. state key laboratory of protein engineering and plant genetic engineering College of Life Sciences Peking University 100871 Beijing China

Using a known GAmyb gene as the probe, two fully identical clones were isolated from a barley aleurone cDNA library. Sequence analysis showed that their 5′ termini are highly homologous to the 3′ termini of GAmyb (97%) and their 3′ termini share no significant homology with any myb genes. Therefore, the deduced protein may hold intact putative GAmyb activation domain but lack the normal DNA binding domain. Northern blot reveals that hvmyb expression in barley aleurone layers is strongly up regulated by gibberellin (GA) and down regulated by abscisic acid (ABA). The tissue and developmental stage specificity of hvmyb was also found, which was only expressed in barley aleurone cells and dropped to non detectable level soon after germination.

关键词： barley aleurone GAmyb GA ABA.

Cloning and Sequencing of a Tomato cDNA of Ethylene-Forming Enzyme

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Chinese Science Bulletin 1994年第5期39卷 430-434页

作者：曹晓风鲁治滨朱玉贤潘乃穟陈章良 state key laboratory of protein engineering and plant genetic engineering Peking UniversityBeijing 100871 PRC

1 Introduction Ethylene, one of the simplest organic molecules with biological activity, is a planthormone that controls many processes of plant growth and development. Its synthesisis strictly regulated in plants. It is induced during some specific developmental stages,such as fruit ripening, seed germination and tissue senescence and abscission.

关键词： ethylene-forming enzyme gene cDNA sequence tomato.

PII,the key regulator of nitrogen metabolism in the cyanobacteria

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Science China(Life Sciences) 2008年第12期51卷 1056-1065页

作者： ZHANG Ying & ZHAO JinDong state key laboratory of protein and plant genetic engineering, College of Life Science, Peking University, Beijing 100871, China 1. state key laboratory of protein and plant genetic engineering College of Life Science Peking University Beijing 100871 China

PII proteins are a protein family important to signal transduction in bacteria and plants. PII plays a critical role in regulation of carbon and nitrogen metabolism in cyanobacteria. Through conformation change and covalent modification, which are regulated by 2-oxoglutarate, PII interacts with different target proteins in response to changes of cellular energy status and carbon and nitrogen sources in cyanobacteria and regulates cellular metabolism. This article reports recent progress in PII research in cyanobacteria and discusses the mechanism of PII regulation of cellular metabolism .

关键词： carbon metabolism cyanobacteria nitrogen metabolism PII protein

Obtaining transgenic rice resistant to rice fungal blast disease by controlled cell death strategy

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Chinese Science Bulletin 2003年第16期48卷 1753-1759页

作者： MAO Shengji, GU Hongya, QU Lijia & CHEN Zhangliang National key laboratory of protein engineering and plant genetic En-gineering, College of Life Sciences, Peking University, Beijing 100871, China Correspondence should be addressed to Qu Lijia (e-mail: qulj@plum. ***) National key laboratory of protein engineering and plant genetic engineering College of Life Sciences Peking University Beijing 100871 China

The strategy of the two-component system, composed of Barnase and Barstar which encode RNase and a specific inhibitor to the RNase respectively, is adopted to obtain transgenic rice resistant to rice fungal blast disease. In this study, two chimeric promoters, induced by rice blast fungus pathogen (Magnaporthe grisea), are fused with Barnase respectively to construct two plant expression vec-tors, pWBNBS and pPBNBS together with the Barstar driven by CaMV 35S promoter. The resistance of the transgenic rice lines to rice blast fungus disease and rice blight disease are evaluated. The results show that (1) the expression of Barnase is induced in rice leaves when inoculated with the spores of Magnaporthe grisea; (2) the induced expression level of Barnase surpasses the level of Barstar, which elicits a similar hypersensitive response (HR) in the leaves, and the transgenic plant shows high resistance to the rice fungal blast disease; and (3) transgenic rice plants also show obvious re-sistance to rice bacterial blight disease. Taken together, these results suggest that the transgenic rice plants harboring this two-component system acquire relatively broad spectrum resistance against pathogens, especially high resistance to rice fungal pathogen.

关键词：转基因水稻水稻病害抗病性受控细胞死亡致病病原体基因工程核糖核酸酶

Infectious in vitro transcripts from cloned cDNA of beet necrotic yellow vein virus RNA3 and RNA4and their functional study

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Science China Chemistry 1995年第4期38卷 438-447页

作者：李毅魏春红田波潘乃穟陈章良 state key laboratory for protein engineering and plant genetic engineering Department of Biology Peking University Beijing 100871 China Institute of Microbiology Chinese Acaderny of SciencesBeijing 100080China

The full-length double-stranded cDNAs of beet necrotie yellow vein virus RNA3 and RNA 4were synthesized by using oligo(dT) 15 as well as RNA3 and RNA4 specific primers, and cloned downstream of the bacteriophage Sp6 RNA polymerase promoter of the transcription vector pGEM3Zf(+). The in vitro "run-off" transcription products obtained in the presence of Sp6 RNA polymerase and template DNA have high biological activities. In the 2 transcription systems, the transcription and capping in 2 separate reactions are more efficient. The simultaneous transcription and capping are not so efficient, but the transcripts are more infectious. Although there are a number of nonviral nucleotide sequences at the 5- and 3’-ends of RNA3 and RNA4 transcripts, the biological activities of both transcripts were not affected. The mechanical coinoculation of sugarbeet roots with the infectious transcripts of the prepared RNA3 and RNA4 and BNYW Rgl isolate has confirmed that RNA3 is the main cause of sugarbeet rhizomania.

关键词： beet necrotic yellow vein virus in vitro transcription function analysis.

Polycomb-group histone methyltransferase CLF is required for proper somatic recombination in Arabidopsis

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Journal of Integrative plant Biology 2014年第6期56卷 550-558页

作者： Na Chen Wang-Bin Zhou Ying-Xiang Wang Ai-Wu Dong Yu Yu state key laboratory of genetic engineering Institute of Plant Biology School of Life Sciences Fudan University

Homologous recombination （HR） is a key process during meiosis in reproductive cells and the DNA damage repair process in somatic cells. Although chromatin structure is thought to be crucial for HR, only a smal number of chromatin modifiers have been studied in HR regulation so far. Here, we investigated the function of CURLY LEAF （CLF）, a Polycomb-group （PcG） gene responsible for histone3 lysine 27 trimethylation （H3K27me3）, in somatic and meiotic HR in Arabidopsis thaliana. Although fluorescent protein reporter assays in pol en and seeds showed that the frequency of meiotic cross-over in the loss-of-function mutant clf-29 was not significantly different from that in wild type, there was a lower frequency of HR in clf-29 than in wild type under normal conditions and under bleomycin treatment. The DNA damage levels were comparable between clf-29 and wild type, even though several DNA damage repair genes （e.g. ATM, BRCA2a, RAD50, RAD51, RAD54,and PARP2） were expressed at lower levels in clf-29. Under bleomycin treatment, the expression levels of DNA repair genes were similar in clf-29 and wild type, thus CLF may also regulate HR via other mechanisms. These findings expand the current knowledge of PcG function and contribute to general interests of epigenetic regulation in genome stability regulation.

关键词： Arabidopsis CLF DNA repair homologous recombination meiosis

Ligand Perception,Activation,and Early Signaling of plant Steroid Receptor Brassinosteroid Insensitive 1

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Journal of Integrative plant Biology 2013年第12期55卷 1198-1211页

作者： Jianjun JiangWang Chi ZhangWang Xuelu Wang state key laboratory of genetic engineering and Institute of plant Biology School of Life SciencesFudan University

Leucine-rich repeat receptor-like kinases （LRR-RLKs） belong to a large group of cell surface proteins involved in many aspects of plant development and environmental responses in both monocots and dicots. Brassinosteroid insensitive 1 （BRI1）, a member of the LRR X subfamily, was first identified through several forward genetic screenings for mutants insensitive to brassinosteroids （BRs）, which are a class of plant-specific steroid hormones. Since its identification, BRI1 and its homologs had been proved as receptors perceiving BRs and initiating BR signaling. The co-receptor BRIl-associated kinase 1 and its homologs, and other BRI1 interacting proteins such as its inhibitor BRI1 kinase inhibitor I （BKI1） were identified by genetic andbiochemical approaches. The detailed mechanisms of BR perception by BRI1 and the activation of BRI1 receptor complex have also been elucidated. Moreover, several mechanisms for termination of the activated BRI1 signaling were also discovered. In this review, we will focus on the recent advances on the mechanism of BRI1 phosphorylation and activation, the regulation of its receptor complex, the structure basis of BRI1 ectodomain and BR recognition, its direct substrates, and the termination of the activated BRI1 receptor complex.

关键词： Brassinosteroid insensitive 1-associated kinase 1 brassinosteroid insensitive 1 kinase inhibitor 1 brassinosteroids brassinosteroidinsensitive 1 phosphorylation receptor-like kinase recognition signaling.

Site-1 protease cleavage site is important for the ER stress-induced activation of membrane-associated transcription factor bZIP28 in Arabidopsis

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Science China(Life Sciences) 2015年第3期58卷 270-275页

作者： SUN Le ZHANG Shuang-Shuang LU Sun-Jie LIU Jian-Xiang state key laboratory of genetic engineering Institute of Plant Biology School of Life Sciences Fudan University

Many sources of stress cause accumulation of unfolded or misfolded proteins in endoplasmic reticulum(ER), which elicits the unfolded protein response(UPR) to either promote cell survival or programmed cell death depending on different developmental context or stress severity. The Arabidopsis membrane-associated transcription factor, b ZIP28, is the functional equivalent of mammalian ATF6, which relocates from the ER to the Golgi where it is proteolytically processed and released from the membrane to the nucleus to mediate the UPR. Although the canonical site-1 protease(S1P) cleavage site on the ER lumen-facing domain is well conserved between b ZIP28 and ATF6, the importance of S1 P cleavage on b ZIP28 has not been experimentally demonstrated. Here we provide genetic evidence that the RRIL573 site, but not the RVLM373 site, on the lumen-facing domain of bZ IP28 is critical for the biological function of b ZIP28 under ER stress condition. Further biochemistry and cell biology studies demonstrated that the RRIL573 site, but not the RVLM373 site, is required for proteolytic processing and nuclear relocation of b ZIP28 in response to ER stress. Our results reveal that S1 P cleavage site plays a pivotal role in activation and function of b ZIP28 during UPR in plants.

关键词： membrane-associated transcription factor ER stress unfolded protein response bZ IP28 S1P S2P