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Construction of single-chromosome DNA library from Lilium regale Wilson

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Chinese Science Bulletin 1998年第5期43卷 434-439页

作者： Benyuan Dang zanmin hu Yihua Zhou Lihua Cui Lanlan Wang Liangcai Li Zhenghua Chen Chinese Acad Sci Inst Genet Beijing 100101 Peoples R China

A protocol of simple rapid microdissection of single-chromosome, amplification and cloning of its DNA from Lilium regale Wilson is described. Single-chromosome, microdissected by micromanipulator, was put into a 0 5 mL Eppendorf tube and digested with Sau3A, and then the Sau3A linker adaptors were ligated to the ends of DNA fragments. After 2 rounds of PCR amplification with one chain of linker adaptor as primer, the PCR products thus obtained have a length of 300-2 500 base pairs (bp) with predominant fragments at about 1 000 bp. Southern blot analysis confirmed that the PCR products originated from the genome of Lilium regale Wilson. By cloning the amplification products from the second round of PCR, single-chromosome DNA library was constructed, in which about as many as 100 000 recombinant clones were produced. A total number of 84 clones were analysed, and it was revealed that the inserts ranged in size from 300 to 1 800 bp, with an average of 780 bp. Compared with the methods described in other literature, this protocol, eliminating the need for enzymatic digestion and ligating micromanipulation of chromosomal DNA in nanoliter volumes, permits the efficient amplification of single chromosome (not tens of chromosomes as reported before) and the fragments (780 bp in average) cloned in this study are longer than those reported before (650 bp in average).

关键词： lilium regale wilson single=chromosome dna library.

Multiple transgenes Populus xeuramericana 'Guariento' plants obtained by biolistic bombardment

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Chinese Science Bulletin 2007年第2期52卷 224-230页

作者： Wang JianGe Su Xiaohua Ji LiLi Zhang BingYu hu zanmin huang RongFeng Tian YingChuan Research Institute of Forestry Chinese Academy of Forestry Beijing 100091 China College of Landscape and art Jiangxi Agricultural University Nanchang 330045 China Institute of Genetics and developmental biology Chinese Academy of Sciences Beijing 100101 China Biotechnology Research Institute Chinese Academy of Agricultural Science Beijing 100081 China Microbiology Institute Chinese Academy of Sciences Beijing 100080 China

A JERF36 regulation gene, a selection marker gene (NPT-II), and the foreign genes levansucrase (SacB), Vitreoscilla hemoglobin (vgb), and Binary coleopterus insect resistance (BtCry3A+OC-I) were co-transferred into Populus xeuramericana 'Guariento' using biolistic bombardment; 25 kanamycin resistant plants were obtained. The results of PCR and Southern hybridization showed that the foreign genes had been integrated into the genome of P. xeuramericana 'Guariento' and 5 genes were all transferred into 7 poplar plants. The results of a BtCry3A ELISA experiment indicated that the BtCry3A gene was expressed in the 7 transgenic poplar plants, and these plants grew well on coastal saline land.

关键词：森林生态系统全球变暖二氧化碳植被

Chromosome microdissection by laser microbeam, chromosomal fragment isolation and amplification in vitro in barley (Hordeum vulgare L.)

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Chinese Science Bulletin 1998年第10期43卷 851-855页

作者： huai Wang Yihua Zhou Benyuan Dang zanmin hu Lanlan Wang Liangcai Li Zhenghua Chen Acad Sinica Inst Genet Beijing 100101 Peoples R China

A method for microdissection, isolation and amplification of plant chromosomal fragments using laser microbeam and a glass microneedle was established. Firstly, 7H chromosome of barley (Hordeum vulgare L.) was dissected by Nd: YAG laserbeam with suitable parameters and the fragment comprising a satellite was isolated with a glass microneedle which was fixed on a micromanipulator. Then, the chromosomal fragment DNA was amplified by LA_PCR (linker adaptor PCR) for two rounds. The size of the DNA fragments of PCR products varied from 500-3 000 bp and the PCR products originated from the genome of barley were verified by Southern hybridization. Compared with previous reports, there are some advantages in this research. The performance is easier, the dissection is more precise and the cost is low. It also permits efficient amplification with only one single chromosome fragment. Laser microbeam_glass microneedle method may be useful in the microdissection of special chromosome regions, especially in plants with middle or small chromosomes.

关键词： Hordeum vulgare L. chromosome microdissection laser microbeam LA_PCR.

Newly Identified Essential Amino Acids Affecting Chlorella ellipsoidea DGAT1 Function Revealed by Site-Directed Mutagenesis

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Newly Identified Essential Amino Acids Affecting Chlorella e...

A mutated rabbit defensinNP-1 produced by Chlorella ellipsoidea can improve the growth performance of broiler chickens

中国生物工程学会第十三届学术年会暨2019年全国生物技术大会

作者： Baocheng Sun Xuejie Guo Chengming Fan Yuhong Chen Jingqiao Wang zanmin hu State Key Laboratory of Plant Cell and Chromosome Engineering Institute of Genetics and Developmental Biology Chinese Academy of Sciences College of Life Sciences University of Chinese Academy of Sciences Institute of Economical Crops Yunnan Agricultural Academy

Diacylglycerol acyltransferase(DGAT) is a rate-limiting enzyme in the synthesis of triacylglycerol(TAG), the most important form of energy storage in plants. Some residues have previously been proven to be crucial for DGAT1 activity. In this study, we used site-directed mutagenesis of the CeDGAT1 gene from Chlorella ellipsoidea to alter 16 amino acids to investigate effects on DGAT1 function. Of the 16 residues(L482 R, E542 R, Y553 A, G577 R, R579 D, Y582 R, R596 D,H603 D, H609 D, A624 R, F629 R, S632 A, W650 R, A651 R, Q658 H, and P660 R), we newly identified 5(L482, R579, H603, A651, and P660) as being essential for DGAT1 function and 7(E542, G577, R596,H609, A624, S632, and Q658) that significantly affect DGAT1 function to different degrees, as revealed by heterologous expression of the mutants in yeast strain INVSc1. Importantly, compared with Ce DGAT1, expression of the mutant CeDGAT1 Y553 A significantly increased the total fatty acid and TAG contents of INVSc1. Comparison among CeDGAT1 Y553 A, Gm DGAT1 Y341 A, At DGAT1 Y364 A,BnDGAT1 Y347 A, and Bo DGAT1 Y352 A, in which tyrosine at the position corresponding to the 553 rd residue in CeDGAT1 is changed into alanine, indicated that the impact of changing Y to A at position 553 is specific for CeDGAT1. Overall, the results provide novel insight into the structure and function of DGAT1, as well as a mutant gene with high potential for lipid improvement in microalgae and plants.

关键词： diacylglycerol acyltransferase site-directed mutagenesis heterologous expression total fatty acid content triacylglycerol

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A mutated rabbit defensinNP-1 produced by Chlorella ellipsoi...

细胞壁重构关键酶木葡聚糖内转糖苷酶/水解酶(XTH)的研究进展

中国生物工程学会第十三届学术年会暨2019年全国生物技术大会

作者： Chengming Fan Jihua Wu Ling Xu Lili Bai Heming Yang Congjuan Yan Qing Wu Yuhong Chen zanmin hu State Key Laboratory of Plant Cell and Chromosome Engineering The Innovative Academy of Seed Design Institute of Genetics and Developmental Biology Chinese Academy of Sciences The 306th Hospital of PLA School of Food andBiological Engineering Jiangsu University Polo Biology Science Park Co. Ltd. College of Life Sciences University of Chinese Academy of Sciences

The demand for alternatives to antibiotics to improve the growth performance of food animals is increasing. Defensins constitute the first line of defence against pathogens in the innate immune system of animals and humans. A transgenic Chlorella ellipsoidea strain producing mN P-1(a mutated rabbit defensin NP-1) was previously obtained in our laboratory. In this study, a process for producing the transgenic strain on a large scale was developed, and the C. ellipsoidea strain producing mNP-1 was used as a feed additive to improve the health and growth performance of chickens. The volume of *** producing mNP-1 can be scaled up to 10,000 L with approximately 100 g/L dry biomass, and the mN P-1 content of transgenic microalgal powder(TMP) was 90–105 mg/L. A TMP-to-regular feed ratio of 1‰, as the optimal effective dose, can promote the growth of broiler chickens by increasing weight by 9.27–12.95%. mNP-1 can improve duodenum morphology by promoting long and thin villi and affect the microbial community of the duodenum by increasing the diversity and abundance of beneficial microbes. These results suggested that transgenic Chlorella producing mNP-1 can be industrially produced and used as an effective feed additive and an alternative to antibiotics for improving the health and growth performance of broiler chickens or other types of food animals/poultry.

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农业生物技术学报 2010年第3期18卷 604-609页

作者：杜丽萍沈昕陈少良胡赞民北京林业大学生物学院教育部树木花卉遗传育种重点实验室中国科学院遗传与发育生物学研究所

细胞壁是目前植物分子生物学研究的热点之一。木葡聚糖(XG)是双子叶植物初生细胞壁中最主要的半纤维素。木葡聚糖内转糖苷酶/水解酶(XTH)是一类催化木葡聚糖分子转移或水解的酶,它能够内切木葡聚糖聚合体,将产生的还原性末端连接到另外... 详细信息

细胞壁是目前植物分子生物学研究的热点之一。木葡聚糖(XG)是双子叶植物初生细胞壁中最主要的半纤维素。木葡聚糖内转糖苷酶/水解酶(XTH)是一类催化木葡聚糖分子转移或水解的酶,它能够内切木葡聚糖聚合体,将产生的还原性末端连接到另外一个木葡聚糖链或水分子上,在细胞壁重构、影响植物生长发育方面有重要作用。本文主要综述了XTH结构﹑功能和影响其功能的因素,概括了目前研究中存在的问题和未来研究方向。

关键词：细胞壁木葡聚糖内转糖苷酶/水解酶(XTH) 结构与功能

OFDM系统中IFFT/FFT处理器的设计与实现

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计算机工程与应用 2011年第1期47卷 60-63页

作者：刘洪涛杨红官胡赞民湖南大学物理与微电子科学学院长沙410082

提出了Radix-4 FFT的优化算法,采用该优化算法设计了64点流水线IFFT/FFT处理器,该处理器可以在64个时钟周期内仅采用3个复数乘法器获得64点处理结果,提高了运算速度,节约了硬件资源。通过Xilinx XC2S300E Spartan2E系列的xc2s300e器件... 详细信息

提出了Radix-4 FFT的优化算法,采用该优化算法设计了64点流水线IFFT/FFT处理器,该处理器可以在64个时钟周期内仅采用3个复数乘法器获得64点处理结果,提高了运算速度,节约了硬件资源。通过Xilinx XC2S300E Spartan2E系列的xc2s300e器件进行下载验证,仿真结果与MATLAB计算结果误差小于0.5%,该处理器已经成功应用于某OFDM通信系统中。

关键词：正交频分复用(OFDM) 反快速傅里叶变换/快速傅里叶变换(IFFT/FFT) 基4算法现场可编程门阵列(FPGA)

Nd:YAG激光微束切割大麦染色体及染色体片段的分离

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中国激光 1998年第7期25卷 657-660页

作者：王槐宋桂英王兰岚胡赞民陈正华中国科学院遗传研究所

以适当参数的ＮｄＹＡＧ激光微束切割大麦７Ｈ染色体后再利用微细玻璃针挑取了７ＨＳ端部片段并放入Ｅｐｐｅｎｄｏｒｆ管中，建立了一种激光微束与玻璃针结合使用微切割、微分离植物染色体片段的方法。

关键词：大麦染色体散切割钕:YAG激光微束

甘蓝型油菜遗传图谱构建及重要性状遗传解析的研究进展

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分子植物育种 2016年第1期14卷 98-116页

作者：李翔范成明卢龙斗胡赞民河南师范大学生命科学学院新乡453002 中国科学院遗传与发育生物学研究所北京100101

油菜是重要的油料作物,甘蓝型油菜在中国油菜生产中占有主导地位。遗传图谱构建是遗传学研究中的重要环节,是从分子水平深入认识和解析复杂农艺性状的基础,也是重要基因图位克隆的手段。自从1991年第一张甘蓝型油菜分子标记连锁图建立以... 详细信息

油菜是重要的油料作物,甘蓝型油菜在中国油菜生产中占有主导地位。遗传图谱构建是遗传学研究中的重要环节,是从分子水平深入认识和解析复杂农艺性状的基础,也是重要基因图位克隆的手段。自从1991年第一张甘蓝型油菜分子标记连锁图建立以来,已构建了多张甘蓝型油菜高密度遗传连锁图谱,并对多个复杂农艺性状进行了解析。本文综述了二十多年来在甘蓝型油菜遗传图谱构建及其重要农艺性状解析领域的研究进展,旨在对甘蓝型油菜的分子改良提供参考。

关键词：甘蓝型油菜遗传图谱分子标记数量性状位点