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Molecular Systematic Studies on Chinese Mandarina Silkworm (Bombyx mandarina M.) and Domestic Silkworm (Bombyx mori L.)

中国生物化学与分子生物学会农业生物化学与分子生物学分会第八次学术研讨会

作者： Yaozhou Zhang~* Qingyou Xia~+ Jie Xu~* Lingying Kong~* Jian Chen~* Zuoming Nie~* Dan Wang~* Jianqing Chen~* Qingliang Zheng~* Qing Chen~* Xiaoyuan Ren~* Jiang Wang~* Zhengbing Lv~* Caiying Jiang~* Lili Liu~* Qing Sheng Yongfeng Jin~(**) Xiangfu Wu~* ~*The key laboratory of Bioreactor and Biopharmacy of Zhejiang Province Institute of Biochemistry,Zhejiang Sci-Tech university,Hangzhou 310018,china ~+The key sericultural laboratory of Agricultural ministry the key laboratory for sericultural sciences and genomics of the ministry of education,college of biotechnology,southwest university,beibei,chongqing 400715,china ~(**)college of Life sciences,Zhejiang university,Hangzhou 310029,china

Mass spectrometry (MS) is an effective method for large-scale proteomic identification and verification of protein-encoding genes present in a *** this study,peptide sequences identified by MS were matched with those from an ORF library of the silkworm (Bombyx mori) genome and a cDNA library to validate putative *** databases were *** first was based on a 6x draft sequence of the silkworm genome and contained 14,632 putative *** second was based on a *** pupal cDNA library that had 3,187 putative proteins,each composed of at least 30 amino acid *** this study,81,000 peptide sequences that had a threshold score of 60% were generated by MS/MS analysis,and 55,400 were chosen for sequence *** searching the two databases,6,649 and 250 protein were matches,and these accounted for approximately 45.4% and 7.8% *** analysis and cataloging of the identified proteins,which were carried out by using several bioinformatic tools,suggested that the matches included proteins with predicted transmembrane domains (TMDs;1,393) and preproteins with a signal peptide (SP,976).These results provide a fundamental understanding of gene expression and function analysis of silkworm proteins.

关键词： Bombyx mori pupae protein identification function analysis ORF genome proteomics

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Agricultural sciences in china 2002年第3期1卷 349-358页

作者： LU Cheng, YU Hong-shi and XIANG Zhong-huai( The key sericultural laboratory of Agricultural ministry, college of Silkworm and Silk engineering , southwest Agricultural university, chongqing 400716) The key sericultural laboratory of Agricultural ministry College of Silkworm and Silk engineering Southwest Agricultural University Chongqing 400716

Molecular systematic studies on mandarina silkworm (Bombyx mandarina M.) in 11 regions in china and 25 representative strains of domestic silkworm (Bombyx mori L.) were conducted using molecular biology techniques. Results obtained from the analysis of DNA polymorphism and clustering of all the silkworm samples provide new evidence for the view that the domestic silkworm originated from the Chinese mandarina silkworm. On the basis of literature reviewing, a new hypothesis on the origin of the domestic silkworm was put forward. It was thought that the domestic silkworm was most probably domesticated from the Chinese mandarina silkworm of different ecotypes including monovoltinism, bivoltinism and multivoltinism; and that the domestic silkworm had the genetic background of monovoltinism, bivoltinism and multivoltinism at the very beginning of the domestication. The current strains of the domestic silkworm of different voltinism are the evolutionary results of thousands of years of rearing and artificial selections.

关键词： Bombyx mandarina M. Bombyx mori L. DNA polymorphism Systematic evolution

Studies on the Mutant Systems of the Bombyx mori Gene Bank

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Agricultural sciences in china 2002年第11期1卷 1277-1285页

作者： LU Cheng, DAI Fang-yin and XIANG Zhong-huai(The key sericultural laboratory, ministry of Agriculture/southwest Agricultural university,chongqing 400716, P.R. china) The key sericultural laboratory Ministry of Agriculture/Southwest Agricultural University Chongqing 400716 P.R.China

Through over ten years of study, more than 1 000 genetic materials including mutant genes, chromosomal variation strains and special genetic materials of Bombyx mori, Linnaeus, collected, introduced or created since 1940s especially late 1980s, have been sorted out and put in order. After identifications and genetic analyses of their morphological, physiological and biochemical characters, the silkworm gene bank was constructed and the preservation system was perfected, and more than 600 silkworm strains were kept in this gene bank. The preserved silkworm mutant genes have covered more than 90% of existent ones across the world, in which, more than 100 are rare and precious mutant genes, and over 60 mutant genes were found and studied for the first time. Through hybrid analyses, linkage tests and three-point gene location tests, a perfect linkage retrieval labeling system of silkworm was established, which included 230 marker genes covering all the 28 linkage groups of Bombyx mori. The gene location system (composite system of recessive genes) of different linkage groups was set up. The intergenic complementation of mutant egg color and third type of maternal heredity egg color have been found, and indicated that the epistatic effect of mutant gene of white egg is universal. Twenty eight independent near isogenic lines marked with morphological mutation gene have been created and a series of novel breeding materials possessing great potential for application such as high feeding efficiency, special sex markers, natural colored silk, resistance to disease, wider feeding range and adjustable parthenogenesis, etc. , have been developed. The sustainable maintenance and management technique system of silkworm gene resources were well established.

关键词： Bombyx mori Mutation Gene bank Genetic analysis Gene-resources development

Characterization of mitochondrial genome of Chinese wild mulberry silkworm, Bomyx mandarina(Lepidoptera:Bombycidae)

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Science china(Life sciences) 2008年第8期51卷 693-701页

作者： PAN MinHui1, YU QuanYou1, XIA YuLing1, DAI FangYin1, LIU YanQun2, LU Cheng1, ZHANG Ze1 & XIANG ZhongHuai1 1 key sericultural laboratory of Agricultural ministry, southwest university, chongqing 400716, china 2 college of Bioscience and biotechnology, Shenyang Agricultural university, Shenyang 110161, china key sericultural laboratory of Agricultural ministry Southwest University Chongqing China college of Bioscience and biotechnology Shenyang Agricultural University Shenyang China

The complete mitochondrial genome of Chinese Bombyx mandarina(ChBm) was *** cir-cular genome is 15682 bp long, and contains a typical gene complement, order, and arrangement iden-tical to that of Bombyx mori(***) and Japanese Bombyx mandarina(JaBm) except for two addi-tional tRNA-like structures:tRNASer(TGA)-like and tRNAIle(TAT)-*** protein-coding sequences are initi-ated with a typical ATN codon except for the COI gene, which has a 4-bp TTAG putative initiator *** of 13 protein-coding genes(PCGs) have a complete termination codon(all TAA), but the re-maining two genes terminate with incomplete *** tRNAs have the typical clover-leaf structures of mitochondrial tRNAs, with the exception of tRNASer(TGA)-like, with a four stem-and-loop *** length of the A+T-rich region of ChBm is 484 bp, shorter than those of JaBm(747 bp) and ***(494―499 bp).Phylogenetic analysis among ***, ChBm, JaBm, and Antheraea pernyi(Anpe) showed that *** is more closely related to ChBm than *** earliest divergence time estimate for ***-ChBm and ***-JaBm is about 1.08±0.18―1.41±0.24 and 1.53±0.20―2.01±0.26 Mya, *** and JaBm diverged around 1.11±0.16―1.45±0.21 Mya.

关键词： Chinese Bombyx mandarina mtDNA phylogeny origin of Bombyx mori

Generation of pancreatic islet cells from human embryonic stem cells

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Science china(Life sciences) 2009年第7期52卷 615-621页

作者： ZHANG DongHui1,2,JIANG Wei1,SHI Yan2 & DENG HongKui1,2 1 key laboratory of Cell Proliferation and Differentiation of the ministry of education,college of Life sciences,Peking university,Beijing 100871,china 2 laboratory of Chemical genomics,Shenzhen Graduate School of Peking university,Shenzhen 518055,china key laboratory of Cell Proliferation and Differentiation of the ministry of education College of Life Sciences Peking University Beijing China laboratory of Chemical genomics Shenzhen Graduate School of Peking University Shenzhen China

Efficiently obtaining functional pancreatic islet cells derived from human embryonic stem(hES) cells not only provides great potential to solve the shortage of islets sources for type I diabetes cell therapy,but also benefits the study of the development of the human pancreas and diabetes *** 2001,hES cells were reported to have the capacity to generate insulin-producing cells by spontaneous differentiation in *** then,many strategies(such as overexpression of key transcription factors,delivery of key proteins for pancreatic development,co-transplantation of differentiated hES cells along with fetal pancreas,stepwise differentiation by mimicking in vivo pancreatic development) have been employed in order to induce the differentiation of pancreatic islet cells from hES ***,patient-specific induced pluripotent stem(iPS) cells can be generated by reprogramming somatic *** cells have characteristics similar to those of ES cells and offer a new cell source for type I diabetes cell therapy that reduces the risk of immunologic *** this review,we summarize the recent progress made in the differentiation of hES and iPS cells into functional pancreatic islet cells and discuss the challenges for their future study.

关键词： human embryonic stem(hES) pancreatic islet cells differentiation induced pluripotent stem(iPS) cells

Ripple-associated high-firing interneurons in the hippocampal CA1 region

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Science china(Life sciences) 2008年第2期51卷 120-126页

作者： WANG Ying, ZHANG Lu, PAN JingWei, XIE Kun, LI ShiQi, WANG ZhiRu & LIN LongNian key laboratory of Brain Functional genomics of ministry of education, East china Normal university, Shanghai 200062, china key laboratory of Brain Functional genomics of ministry of education East China Normal University Shanghai China

By simultaneously recording the activity of individual neurons and field potentials in freely behaving mice, we found two types of interneurons firing at high frequency in the hippocampal CA1 region, which had high correlations with characteristic sharp wave-associated ripple oscillations (100―250 Hz) during slow-wave sleep. The firing of these two types of interneurons highly synchronized with ripple oscillations during slow-wave sleep, with strongly increased firing rates corresponding to individual ripple episodes. Interneuron type I had at most one spike in each sub-ripple cycle of ripple episodes and the peak firing rate was 310±33.17 Hz. Interneuron type II had one or two spikes in each sub-ripple cycle and the peak firing rate was 410±47.61 Hz. During active exploration, their firing was phase locked to theta oscillations with the highest probability at the trough of theta wave. Both two types of interneurons increased transiently their firing rates responding to the startling shake stimuli. The results showed that these two types of high-frequency interneurons in the hippocampal CA1 region were involved in the modulation of the hippocampal neural network during different states.

关键词： hippocampus ripple oscillation theta rhythm interneuron

Microarray analysis of gene expression profiles in the bovine mammary gland during lactation

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Science china(Life sciences) 2010年第2期53卷 248-256页

作者： HOU XiaoMing,LI QingZhang & HUANG TianYing key laboratory of Dairy Science of education ministry,Northeast Agricultural university,Harbin 150030,china key laboratory of Dairy Science of education ministry Northeast Agricultural University

Mammary glands undergo functional and metabolic changes during virgin,lactation and dry periods.A total of 122 genes were identified as differentially expressed,including 79 up-regulated and 43 down-regulated genes during lactation compared with virgin and dry *** ontology analysis showed the functional classification of the up-regulated genes in lactation,including transport,biosynthetic process,signal transduction,catalytic activity,immune system process,cell death,and positive regulation of the developmental *** data clarified molecular events in bovine mammary gland lactation.

关键词： mammary gland microarray lactation genomics gene expression profile

Cloning, Expression, Purification and Crystallization of NHR3 Domain from Acute Myelogenous Leukemia-related Protein AML1-ETO

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 566-570页

作者： Hai-Tao YANG , Dong-Hui WU1 , Xiao-Yu XUE, Wen-Xue LIANG1, Xiao-Yu MIAO,Hai PANG , and Sai-Juan CHEN1laboratory of Structural Biology, Department of Biological sciences and biotechnology & Protein sciences laboratory of MOE, Tsinghua university, Beijing 100084, china Shanghai Institute of Hematology, State key laboratory of Medical genomics,1 Shanghai Second Medical university, Shanghai 200023, china laboratory of Structural Biology Department of Biological Sciences and Biotechnology & Protein Sciences Laboratory of MOE Tsinghua UniversityBeijing 100084 China Shanghai Institute of Hematology State Key Laboratory of Medical Genomics Shanghai Second Medical UniversityShanghai 200023 China

The t(8;21) translocation is one of the most frequent chromosome abnormalities in acute myeloid leukemia. This translocation creates a fusion between the acute myelogenous leukemia 1 (AML1, a transcription factor) gene on chromosome 21 and the eight-twenty-one (ETO, a zinc finger nuclear protein) gene on chromosome 8, leading to the repression of certain AML1 target genes. We cloned NHR3 domain coding fragment into vector pGEX-6p-1 using PCR and obtained recombinant plasmid pGEX-6p-1-NHR3, which can be induced to stably overexpress fusion protein in E. coli. Through the purification on GST affinity chromatography column and PreScission protease cleavage, a large amount of NHR3 protein with high purity was obtained. In order to avoid possible interference of some strong negative charged molecules, NHR3 protein was further purified by Mono Q anion exchange chromatography. The NHR3 crystals were grown with hanging drop/vapor diffusion method and the first crystals appeared after four weeks at 18 °C in 0.2 M Tris-sodium citrate dihydrate, 0.1 M sodium cacodylate, pH 6.5, and 30% iso-propanol (V/V). ESI mass spectrum showed that the molecular weight of this domain was in agreement with its primary structure sequence prediction, and circular dichroism spectral data (190–250 nm) showed that NHR3 had a well-defined secondary structure of 25.9% α-helix, 23.2% random coil and 50.9% turn, which was consistent with GOV4 software prediction.

关键词： AML1-ETO NHR3 crystallization ESI mass spectrum circular dichroism spectrum glutathione S-transferase (GST)

Antisense expression of a rice cellular apoptosis susceptibility gene (OsCAS} alters the height of transgenic rice

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Progress in Natural Science:Materials International 2007年第1期17卷 39-43页

作者： XU Chunxiao and HE Chaozu ( National key laboratory of Plant genomics, Institute of Microbiology, Chinese Academy of sciences, Beijing 100080, china Graduate university of Chinese Academy of sciences, Beijing 100039, china Shenzhen laboratory of Superhybrid Rice Research, Graduate School at Shenzhen, Tsinghua university, Shenzhen 518055, china) National key laboratory of Plant genomics Institute of Microbiology Chinese Academy of Sciences Beijing 100080 China

Cellular apoptosis susceptibility (CAS) gene plays important roles in mitosis, development and export of importin a from the nucleus, but its function in plant is unknown. In this study, a rice CAS ortholog (OsCAS), which encodes a predicted protein of 983 amino acids with 62% similarity to human CAS, was identified. DNA gel blot analysis revealed a single copy of OsCAS in the rice genome. A 973 bp fragment at the 3' end of OsCAS cDNA was cloned from rice cDNA library and transferred into rice in the antisense direction under the control of CaMV 35S promoter via Agrobacterium-mediated transformation method, 105 transgenic lines were obtained. Expression of OsCAS was suppressed in the antisense transgenic lines as revealed by semi-quantitative RT-PCR. The antisense transgenic lines showed dwarf phenotypes. The results indicated that OsCAS was involved in culm development of rice.

关键词： antisense dwarf ortholog OsCAS rice.