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Seasonal influence on the development of parthenotes and cloned embryos in pigs

Progress in Natural Science:Materials International 2007年第10期17卷 1172-1176页

作者： Zhang Kun1, Zhang Yunhai3,4, Pan Dengke5, Ma Yufang1, Wei Hengxi1, Zhang Jian6. Li Xuyang7, Wang Shaohua1, Li Yan2, Dai Yunping1,2 and Li Ning1(1. State Key Laboratory for Agrobiotechnology, College of Biological Science, china agricultural university, beijing 100094. china 2. JiPulin Biotech Ltd., beijing 100094. china 3. Danish Institute of agricultural Sciences, 8830 Tjele, Denmark 4. College of animal Science and Technology, Anhui agricultural {6., Hefei 230036. china 5. Institute of animal Sciences, Chinese Academy of agricultural Sciences, beijing 100094. china 6. animal clinical hospital, china agricultural university, beijing 100094. china 7. Hebei Minghui Group of Pigs Breeding, Sanhe 06.200, china) State Key Laboratory for Agrobiotechnology College of Biological Science China Agricultural University Beijing 100094 China Danish Institute of agricultural Sciences 8830 Tjele Denmark College of animal Science and Technology Anhui Agricultural University Hefei 230036 China Institute of animal Sciences Chinese Academy of Agricultural Sciences Beijing 100094 China animal clinical hospital China Agricultural University Beijing 100094 China Hebei Minghui Group of Pigs Breeding Sanhe 065200 China JiPulin Biotech Ltd. Beijing 100094 China

Seasonal influence on the developmental ability of porcine embryos was studied by comparisons of the number of cumulus-oocytes-complexes (COCs) per ovary pair and meiotic maturation ability of oocytes in different seasons; and by the observations on developmental competence of the parthenogenetic oocytes and cloned embryos collected from different seasons. We found that the number of COCs per ovary pairs was significantly higher in spring compared to the other seasons (P<0.05). However, no significant difference was found in the rate of oocytes progressing to MⅡ in four seasons (P<0.05). In addition, the rate of blastocyst formation of parthenogenetic embryos in summer was obviously declined compared to the other seasons (P<0.05), and an increased blastocyst rate of cloned porcine embryos was found in spring compared to autumn and winter (P<0.05). The results suggest that there should be a seasonal influence on the developmental competence of porcine embryos.

关键词： pig cloned parthenogenetic season

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Genetic analysis on 3'- terminal flanking region of uncoupling protein 3 in different pig breeds

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Chinese Science Bulletin 2002年第18期47卷 1541-1543页

作者： FANG Meiying, ZHAO Xingbo, LI Ning & WU ChangxinCollege of animal Science and Technology, china agricultural university, beijing 100094. china State Key Laboratory for Agrobiotechnology, china agricultural university, beijing 100094. china College of animal Science and Technology China Agricultural University Beijing China State Key Laboratory for Agrobiotechnology China Agricultural University Beijing China

The 3’-terminal flanking region of porcine uncoupling protein 3 (UCP3) was cloned, the sequence data revealed 15 nucleotide substitutions among Landrace and three Chinese native pig breeds named Neijiang, Minpig and Erhualian. The continuous 9 polymorphic sites were checked by PCR-RFLP, the results indicated that Erhualian had extraordinary gene frequency, presented most significant difference by x2 test compared with Landrace, Largewhite, Neijiang and Minpig respectively, significant level compared with Meishan; and Meishan also had significant difference compared with Landrace and Minpig respectively. These results can be concluded that Taihu pigs have special genetic characteristics among pig breeds.

关键词： pig uncoupling protein gene clone PCR-RFLP.

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Aberrant histone H4 acetylation in dead somatic cell-cloned calves

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Progress in Natural Science:Materials International 2008年第7期18卷 813-818页

作者： Lei Zhang a,b,*, Shaohua Wang a, Qiang Li a, Xiangdong Ding c, Yunping Dai a, Ning Li a,* a State Key Laboratory for Agrobiotechnology, china agricultural university, beijing 100094. china b beijing Gene & Protein Biotechnology Co. LTD, beijing 100094. china c College of animal Science and Technology, china agricultural university, beijing 100094. china State Key Laboratory for Agrobiotechnology China Agricultural University Beijing 100094 China beijing Gene & Protein Biotechnology Co. LTD Beijing 100094 China College of animal Science and Technology China Agricultural University Beijing 100094 China

In somatic cell-cloned animals, inefficient epigenetic reprogramming can result in an inappropriate gene expression and histone H4 acetylation is one of the key epigenetic modifications regulating gene expression. In this study, we investigated the levels of histone H4 acetylation of 11 development-related genes and expression levels of 19 genes in lungs of three normal control calves and nine aber- rant somatic cell-cloned calves. The results showed that nine studied genes had decreased acetylation levels in aberrant clones (p 0.05). Whereas 13 genes had significantly decreased expression (p 0.05), and only one gene had higher expression level in clones (p < 0.05). Furthermore, FGFR, GHR, HGFR and IGF1 genes showed lowered levels of both histone H4 acetylation and expression in aberrant clones than in controls, and the level of histone H4 acetylation was even more lowered in aberrant clones than those in controls. It was suggested that the lower levels of histone H4 acetylation in aberrant clones caused by the previous memory of cell differentiation might not support enough chromatin reprogramming, thus affecting appropriate gene expressions, and growth and development of the cloned calves. To our knowledge, this is the first study on how histone H4 acetylation affects gene expression in organs of somatic cell-cloned calves.

关键词： Somatic cell-cloned calves Histone H4 acetylation Gene expression Lungs

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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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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 {6.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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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 {6. university, Kunming 6.0201, 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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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(P6. σP,*** next introgressed a fragment via the y0 marker into a wild stock background over 20 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(P<0.0001).This indicated that there must be a single QTL for low body weight that is tightly linked to the y0 *** then shortened the introgressed fragment to less than 1.5 cM by a deeper MAI using the y0 marker and the white *** narrower fragment also resulted in a similar decrease in body weight to that induced by y0(OR)20,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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Relationship Between Agriculture and Ecological Deterioration,Restoration and Reconstruction in Loess Plateau Areas of Northwest china

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agricultural Sciences in china 2002年第1期1卷 114-120页

作者： CHENG Xu( china agricultural university, beijing 100094. china agricultural university Beijing 100094

A comprehensive analysis was conducted on the current situation as well as historic succession processor agriculture and ecological conditions in Northwest china, in particular in the Loess Plateau, by means of a inter-disciplinary approach of ecology, agro-geohistory, and agronomy. It was concluded that the fundamental causes responsible for the ecological deterioration, signed by the poor agro-productivity and serious soil erosion, had been the extensive cropping system by ever-increasing reclamation on semi-drought slope land, where initiated a natural vegetation of brushy grasses with sparse trees, thus suitable only for animal husbandry. Based on an identification of specific actual status of china, several countermeasures of correctly dealing with the relationship between agricultural development and the ecological restoration and reconstruction were proposed, including to get breakthrough first in the transection area by means of the rational use of resources and the raising of agro-system productivity, to promoting 'water conservancy-type ecological agriculture' through the radical shift of traditional production pattern, and the adoption of a policy of proper degree of grain self-reliance, as well as to construct livestock industry base in Northwest china.

关键词： Loess Plateau Area of Northwest china agricultural ecology

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Production of porcine cloned transgenic embryos expressing green fluorescent protein by somatic cell nuclear transfer

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Science china(Life Sciences) 2006年第2期49卷 164-171页

作者： ZHANG Yunhai1, PAN Dengke1,2, SUN Xiuzhu1, SUN Guojie1, WANG Xiaobo1, LIU Xiaohui1, LI Yan1, DAI Yunping1 & LI Ning1 1. State Key Laboratory for Agrobiotechnology, china agricultural university, beijing 100094. china 2. College of animal Science and Technology, china agricultural university, beijing 100094. china State Key Laboratory for Agrobiotechnology China Agricultural University Beijing China College of animal Science and Technology China Agricultural University Beijing China

In the present study, nuclear transferred embryos (NTEs) were reconstructed by using pig fetal fibroblasts as donors; in vitro matured oocytes as recipients. The effects of G418 selection on donor cells, duration of IVM of prepubertal gilt oocytes; oxygen tension in IVM of oocytes were investigated. The results were as follows: (i) When G418 selected cells expressing GFP were used as donors, the cleavage rate of NTEs decreased drastically in comparison to NTEs derived from donors without antibiotic selection (47.5% vs. 71.6., p>0.05). For the blastocyst rate, no significant difference was observed between two groups (10% vs. 10.4%, p6.to 42 h (83.6. vs. 96.7%, p>0.05). However, no statistical difference was observed between NTEs derived from oocytes of 36.h IVM group; NTEs from oocytes of 42 h IVM group in the rates of cleavage (59.3% vs. 73.6., p6. vs. 6..1%, p6.to 42 h, while no significant differences were observed for developmental potential of transgenic embryos. Thus IVM 42 h is the better choice in order to obtain maximum number of MII oocytes as recipients. (iii) Lower oxygen tension; higher oxygen tension in IVM have no significant effect on development of cloned embryos.

关键词： transgenic cloning, GFP, Pig, early development.

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