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Effects of chemical activation and season on birth efficiency of cloned pigs

science china(Life sciences) 2009年第7期52卷 657-664页

作者： MA YuFang1,2,LI Yan3,WEI HengXi1,LI QiuYan3,FANG Rui1,ZHAO Rui3,ZHANG Kun1,XUE Kai1,LOU YanKun1,DAI YunPing3,LIAN LinSheng2 & LI Ning1 1 State Key Laboratory of Agrobiotechnology,college of Biological sciences,china agricultural university,beijing 100193,china 2 college of animal science and technology,china agricultural university,beijing 100193,china 3 beijing Genprotein Biotechnology Company,beijing 100193,china State Key Laboratory of Agrobiotechnology College of Biological Sciences China Agricultural University Beijing China college of animal science and technology China Agricultural University Beijing China beijing Genprotein Biotechnology Company Beijing China

The effects of chemical activation on birth efficiency of cloned pigs were studied by investigating the developmental process from porcine oocyte activation to birth of cloned *** different activation methods were used:(i) Electroporation(Ele);(ii) Ele followed by incubation with 6-dimethylaminopurine(6-DMAP);and(iii) Ele followed by a treatment with cycloheximide(CHX).In experiment 1,the rates of cleavage,developmental rates and cell number of porcine parthenogenetic(PA) embryos were investigated in the three treatment *** experiment 2,NT embryos produced by the three different activation treatments were compared for the rates of cleavage,development and cell ***,the effects of Ele and Ele+CHX activation methods on birth efficiency of cloned pigs were *** activated oocytes treated by combination activation generally showed a higher(P<0.05) blastocyst rate and produced more expanded blastocysts than oocytes activated with *** rates of cleavage and total cell number of parthenotes were not significantly *** embryos activated with 6-DMAP developed into blastocyst and expanded blastocyst stages at a significantly(P<0.05) higher rate than those treated with Ele,but the developmental capability was dramatically decreased in NT *** the CHX activation method,the NT embryo blastocyst rate was substantially(P<0.05) increased although the production of expanded blastocysts was not significantly different from that by the other two *** birth rate of cloned pigs increased in the CHX group,though the rate was not significantly different from *** effects of season on developmental rate of the porcine PA embryos and birth rate of cloned pigs were also examined in our *** oocytes collected in the spring had higher developmental capabilities than those collected in the ***,no difference in birth rate of the cloned pigs was found between the oocytes collected in the two *** re

关键词： parthenogenetic activation chemical activation nuclear transfer pig season

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Study on Tibetan Chicken embryonic adaptability to chronic hypoxia by revealing differential gene expression in heart tissue

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science china(Life sciences) 2009年第3期52卷 284-295页

作者： LI Mei & ZHAO ChunJiang college of animal science and technology,china agricultural university,beijing 100193,china college of animal science and technology China Agricultural University Beijing China

Oxygen concentration is essential for appropriate *** can exert a significant impact on physiological alteration of the cell and *** Chicken(Gallus gallus) is a Chinese indigenous breed inhabiting in Tibetan areas,which is also a chicken breed living at high altitude for the longest time in the *** has developed an adaptive mechanism to hypoxia,which is demonstrated by that Tibetan Chicken has much higher hatchability than low-land chicken breeds in high-altitude areas of *** the present study,Tibetan Chicken fertilized full sib eggs were incubated up to Hamburger-Hamilton stage 43 under 13% and 21% oxygen concentration,*** Chicken and Dwarf Recessive White Chicken were used as control *** hearts in all of the 3 chicken breeds under hypoxic and normoxic conditions were isolated and hybridized to Genechip Chicken Genome Array to study molecular mechanisms underlying the adaptation to high altitude of Tibetan *** a result,50 transcripts highly expressed in hypoxia are screened *** up-regulated genes,some are involved in the gene ontology(GO) such as cell growth,cell difference,muscle contraction and signal ***,the expression levels of 21 transcripts are lower in hypoxia than those in *** down-regulated genes take part in cell communication,ion transport,protein amino acid phosphorylation and signal ***,gene enrichment analyses of these differential gene expressions are mainly associated with immune system response and ion channel activity in response to ***,the transcriptional expression profiles analyzed by hierarchical clustering and CPP-SOM software in all of the 3 different chicken breeds revealed that Tibetan Chicken is much closely related to Shouguang Chicken rather than Dwarf Recessive White *** addition,12 transcripts of Tibetan Chicken breed-specific expressed genes were identified,which seem to result in a more

关键词： adaptation chronic hypoxia gene chip chicken genome array Tibetan Chicken(Gallus gallus) differential gene expression

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yellow^0,a marker for low body weight in Drosophila melanogaster

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science china(Life sciences) 2009年第7期52卷 672-682页

作者： Li XinHai & Deng XueMei college of animal science and technology,china agricultural university,beijing 100193,china college of animal science and technology China Agricultural University Beijing China

Marker-assisted selection(MAS) is an important modern breeding technique,but it has been found that the effect of the markers for quantitative trait loci(QTL) is inconsistent,leading in some cases to MAS failure and raising doubts about its *** the model organism Drosophila melanogaster was employed to study whether an effective marker could be found and applied to *** crossed the stock carrying the y0 marker(a recessive mutation allele of the yellow gene on the X chromosome) with three other stocks carrying corresponding wild-type markers in an F2 design,and found that the y0 marker was in significant association with low body weight(P20 generations of marker-assisted introgression(MAI),and constructed the introgression stock y0(OR)20 in which body weight decreased by 13% and 7%,in female and male,respectively,compared to the wild stock(P20,indicating that the QTL for low body weight is located within this less-than-1.5 cM *** characteristics of the y0 marker by PCR amplification and Southern blotting revealed that yellow gene was deficient in the y0 stock,leading to disappearance of melanin from the cuticle and probably influencing the developmental *** above results confirmed the existence of effective QTL markers applicable to MAS breeding schemes,and their potential application in breeding new stocks.

关键词： Drosophila low body weight yellow quantitative trait loci marker-assisted selection introgression

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Neuroglobin mutation associated with hypoxia adaptation in Tibet chicken

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Progress in Natural science:Materials International 2007年第12期17卷 1419-1424页

作者： Zhang Lifan1, Liu Chong2, Lian Linsheng3, Zhao Chunjiang1, Bao Haigang1 and Wu Changxin1(1. college of animal science and technology, china agricultural university, beijing 100094, china 2. college of Biological science, china agricultural university, beijing 100094, china 3. college of animal science and technology, Yunnan agricultural university, Kunming 650201, china) college of animal science and technology China Agricultural University Beijing 100094 China college of Biological science China Agricultural University Beijing 100094 China college of animal science and technology Yunnan Agricultural University Kunming 650201 China

Globin protein family plays an important role in storing and transporting oxygen. As a newly reported globin, the revealed function of neuroglobin includes binding and storing oxygen as well as facilitating the utilization of oxygen in neuronal cells. In the present study, mutations in exons of chicken Ngb gene were identified with the method of sequencing and created restriction site PCR in Tibet chicken and other four lowland chicken breeds. The mutations of Lys-2224(E4)-Asn and Ser-2279(E4)-Gly were identified in exon 4 of the gene. The Lys-2224(E4)-Asn mutation existed only in Tibet chicken and the mutation frequencies increased with increasing altitude. Analysis of the haplotype and diplotype of the two mutations in Tibet chicken populations of different altitudes showed that the frequencies of TG haplotype and TTGG diplotype also increased with increasing altitude, while the reverse tendency was found on GGAA diplotype. Under the hypoxic simulation incubation, the main haplotype was TG in living embryos and GA in dead embryos. The results showed that the Lys-2224(E4)-Asn mutation may be a specific mutation associated with hypoxia adaptation in Tibet chicken.

关键词： Tibet chicken neuroglobin hypoxia adaptation Ngb gene mutation.

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Expression of the Tribolium castaneum (Coleoptera: Tenebrionidae) hsp83 gene and its relation to oogenesis during ovarian maturation

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Journal of Genetics and Genomics 2010年第8期37卷 513-522页

作者： Jingjing Xu 1, Juan Shu 1, Qin Zhang Key Laboratory of animal Genetics and Breeding of the Ministry of Agriculture, college of animal science and technology, china agricultural university, beijing 100193, china Key Laboratory of animal Genetics and Breeding of the Ministry of Agriculture College of Animal Science and Technology China Agricultural University Beijing 100193 China

Heat shock proteins （HSP） can protect organisms and cells from thermal damage. In this study, we cloned the full length cDNA encoding the HSP83 protein （the homologue of HSP90） of Tribolium castaneum（red flour beetle）. The isolated cDNA contains the full coding sequence, a partial 5＇ untranslated region of 55 bp and the complete 3＇ untranslated region. We found the hsp83 gene is located on chromosome 5 of the T. castaneum genome. The predicted HSP83 protein sequence has a high similarity （on average 86.77%） with that of other insect species. The expression of the hsp83 gene in the whole body and in the ovary could be induced with heat stress （40℃ for 1 h） in newly hatched （within 3 h post emergence） and mature （10 days post emergence） beetles. Under normal conditions, the hsp83 expression in the ovary is about 3-fold higher than in the whole body at both stages. No significant difference in hsp83 expression was observed between the two ovarian developmental stages regardless if the beetles were treated with heat shock or not. The expression of the HSP83 protein in the whole body could also be induced with heat stress in newly hatched and mature beetles. However, in the ovary, HSP83 was only expressed in the follicle cells of mature beetles and not in newly hatched beetles, regardless if the beetles were treated with heat shock or not. Furthermore, the females were not able to produce mature oocytes after knock-down of the hsp83 expression by injecting dsRNA. These results suggest that the HSP83 protein is involved in protection against heat stress and could be involved in oogenesis during ovarian maturation of T. castaneum.

关键词： hsp83 cDNA cloning Tribolium castaneum expression heat shock

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Simulation study on the efficiencies of MOET nucleus breeding schemes applying marker assisted selection in dairy cattle

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science china(Life sciences) 2009年第3期52卷 296-306页

作者： LUO WeiZhen,WANG YaChun & ZHANG Yuan Department of animal Genetics & Breeding,college of animal science and technology,china agricultural university,beijing 100094,china Department of animal Genetics & Breeding College of Animal Science and Technology China Agricultural University Beijing China

Advantages of breeding schemes using genetic marker information and/or multiple ovulation and embryo transfer(MOET) technology over the traditional approach were extensively evaluated through *** yield was the trait of interest and QTL was the genetic marker *** dairy cattle breeding scenarios were considered,i.e.,traditional progeny testing breeding scheme(denoted as STANPT),GASPT scheme including a pre-selection of young bulls entering progeny testing based on their own QTL information,MOETPT scheme using MOET technology to generate young bulls and a selection of young bulls limited within the full-sib family,GAMOPT scheme adopting both QTL pre-selection and MOET technology,COMBPT scheme using a mixed linear model which considered QTL genotype instead of the BLUP model in GAMOPT,and three non-progeny testing schemes,*** MOET,GAMO and COMB schemes,corresponding to MOETPT,GAMOPT and COMBPT with progeny testing being part of the *** were selected based on their breeding value which was estimated under an animal model *** selection over 17 years was performed in the simulations and 30 replicates were designed for each *** influences of using QTL information and MOET technology on favorable QTL allele frequency,true breeding values,polygenetic breeding values and the accumulated genetic superiority were extensively evaluated,for five different populations including active sires,lactating cows,bull dams,bull sires,and young *** results showed that the combined schemes significantly outperformed other approaches wherein accumulated true breeding value *** difference between schemes exclusively using QTL information or MOET technology was not *** STANPT scheme was the least efficient among the 8 *** schemes using MOET technology had a higher polygenetic response than others in the 17th *** increases of frequency of the favorable QTL allele varied more greatly across t

关键词： dairy cattle marker-assisted selection two-stage selection MOET QTL

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Theoretical analysis of alloy multiplying factor by the empirical electron theory of solids and molecules

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science china(Technological sciences) 2007年第6期50卷 807-814页

作者： SONG YuePeng1,2, LIU GuoQuan2, LI ZhiLin3, FENG ChengMing1 1 Mechanical and Electronic Engineering college, Shandong agricultural university, Tai’an 271018, china 2 college of Materials science and Engineering, university of science and technology, beijing 100083, china 3 college of Material science and Engineering, beijing university of Chemical technology, beijing 100029, china Mechanical and Electronic Engineering college Shandong Agricultural University Tai’an China college of Materials science and Engineering University of Science and Technology Beijing China college of Material science and Engineering Beijing University of Chemical Technology Beijing China

By introducing the distribution probability of structural units in austenite contain- ing alloying elements and considering its effects on phase transformation, this paper establishes a calculation model of distribution probability of structural units. A new valence electron structure (VES) parameter-transformation effect coefficient of alloying elements (HL), is defined and then studied both theoretically and ex- perimentally. The relationship between the parameter HL and the multiplying factor (the quenching capability factor) of alloying elements is studied. The results indi- cate that the two parameters (HL and the quenching capability factor) have the same variation characteristic and substance feature. Therefore, the multiplying factor virtually expresses the relative quantity of structural units in the alloying elements-containing austenite.

关键词： multiplying factor (the quenching capability factor), empirical electron theory of solids and molecules, transformation effect coefficient of alloying elements, structural units in austenite containing alloying elements

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A five-fold pig bacterial artificial chromosome library:a resource for positional cloning and physical mapping

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Progress in Natural science:Materials International 2006年第8期16卷 889-892页

作者： LIU Wei, ZHANG Ying, LIU Zhaoliang, GUO Li, WANG Xiaobo, FEI Jing, FENG Jidong, ZHAO Rui, HU Xiaoxiang and LI Ning (The State Key Laboratory for Agro-Biotechnology, college of Biology, china agricultural university, beijing 100094, china college of animal science and technology, china agricultural university, beijing 100094, china) The State Key Laboratory for Agro-Biotechnology College of Biology China Agricultural University Beijing 100094 China college of animal science and technology China Agricultural University Beijing 100094 China

A pig BAC library was constructed with genomic DNA from a male Erhualian pig. After partial digestion with HindⅢor BamHⅠthe fragments obtained were cloned into the pBeloBAC11 vector. The library consists of 184320 clones which stored in 480 pieces 384-well plates (20 plates per superpool). A two-step 4-dimension PCR screening system was established to screen the positive clones. An average insert size of 128 kb was estimated from 105 randomly isolated clones, which indicates that the library is more than five times of genomic coverage. For the demonstration of the probability to pick out any unique genes or DNA markers from the library, 10 single-copy genes were screened out and the positive clones were yielded between 1 and 8 with an average of 3.6. Positive superpools were obtained for 32 microsatellite markers selected from different regions of pig genome. The number of positive superpools for each marker varies from 1 to 9 with an average of 4.78. This BAC library provides an additional resource for pig physical mapping and gene identifica-

关键词： pig BAC library PCR screening system.

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Generation of fad2 transgenic mice that produce omega-6 fatty acids

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science china(Life sciences) 2009年第11期52卷 1048-1054页

作者： CHEN Qing1, LIU Qing2, WU ZhiFang1, WANG ZongYi3 & GOU KeMian1 1 State Key Laboratory for Agrobiotechnology, china agricultural university, beijing 100193, china 2 Blackmountain Laboratory, CSIRO Plant Industry, Canberra, Australia 3 college of animal science and technology, china agricultural university, beijing 100193, china State Key Laboratory for Agrobiotechnology China Agricultural University Beijing China Blackmountain Laboratory CSIRO Plant Industry Canberra Australia college of animal science and technology China Agricultural University Beijing China

Fatty acid desaturase-2 (FAD2) introduces a double bond in position 12 in oleic acid (18:1) to form linoleic acid (18:2 n-6) in higher plants and microbes. A new transgenic expression cassette, containing CMV promoter/fad2 cDNA/SV40 polyA, was constructedto produce transgenic mice. Among 63 healthy offspring, 10 founders (15.9%) integrated the cotton fad2 transgene into their genomes, as demonstrated by PCR and Southern blotting analysis. All founder mice were fertile and heterozygous fad2 female and nontransgenic littermates were used for fatty acid analysis using gas chromatography. One fad2 transgenic line showed substantial differences in the fatty acid profiles and the level of linoleic acid was increased 19% (P20:4 n-6) in muscles and liver, compared to their nontransgenic littermates. The results indicate that the plant fad2 gene can be functionally expressed in transgenic mice and may playan active role in conversion of oleic acid into linoleic acid.

关键词： fatty acid desaturase FAD2 transgenic mouse oleic acid linoleic acid arachidonic acid

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Purification of a Novel Antibacterial Short Peptide in Earthworm Eisenia foetida

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Acta Biochimica et Biophysica Sinica 2004年第4期36卷 297-302页

作者： Yan-Qin LIU1,2, Zhen-Jun SUN1*, Chong WANG1, Shi-Jie LI2, and Yu-Zhi LIU2 college of Resource and Environment science, china agricultural university, beijing 100094, china 2college of animal science, Hebei agricultural university, Baoding 071001, china college of Resource and Environment science China Agricultural UniversityBeijing 100094 China college of animal science Hebei Agricultural UniversityBaoding 071001 China

A novel antimicrobial short peptide was purified from earthworm (Eisenia foetida) by a five-step protocol including ammonium sulfate precipitation, ultrafiltration, DE-52 ion exchange chromatography, Sephadex G-10 column chromatography, and C-18 reversed-phase HPLC techniques. The purified peptide was applied to the MALDI-TOF MS to determine the molecular mass and was also subjected to TOF MS-MS analysis to determine the amino acid sequence. As a result, a novel antibacterial peptide, named OEP3121, was obtained, with the molecular mass of 510.8 Da and the sequence being “ACSAG”.

关键词： antibacterial peptide earthworm purification

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