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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.

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 riations (p > 0.05). Whereas 13 genes had significantly decreased expression (p rrant clones, five genes showed no significant differences between controls and clones (p > 0.05), and only one gene had higher expression level in clones (p rthermore, 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

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.

Genetic polymorphisms of milk epithelial mucin of yaks

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progress in Natural Science:Materials International 2006年第11期16卷 1222-1226页

作者： ZHENG Yucai~1, ZHAO Xingbo~2**, JIN Suyu~1, pENG Xianwen~1 and BAI Wenlin~1 (1. College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, china 2. state key laboratory for Agrobiotechnology and Ministry of Agriculture key laboratory of animal Genetics, Breeding and reproduction, china Agricultural University, beijing 100094, china) College of Life Science and Technology Southwest University for Nationalities Chengdu 610041 China state key laboratory for Agrobiotechnology Ministry of Agriculture Key Laboratory of Animal Genetics Breeding and Reproduction China Agricultural University Beijing 100094 China

Milk epithelial mucin (MUC1) of yaks was separated by SDS-polyacrylamide gel electrophoresis. A total of nine types of MUC1 were revealed in 427 yaks from five yak breeds including Maiwa yak, Jiulong yak, Tianzhu white yak, Qinghai yak and Tibetan yak. The molecular weights of MUC1 are from 163 kD to 208 kD, most of which are larger than those of bovine. population genetic analysis shows that the gene frequency and genotype frequency of yak MUC1 differ among breeds, with relatively high gene heterozygosity. The five yak breeds studied fall into two groups according to their milk MUC1 gene frequency, suggesting that milk MUC1 exhibits specificities for local yak groups.

关键词： yak milk genetic polymorphism MUC1.

Aberrant DNA methylation patterns in cultured mouse embryos

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progress in Natural Science:Materials International 2005年第12期15卷 1079-1083页

作者： HOU Jian,CUI Xiuhong,LEI Tinghua,LIU Lei,AN Xiaorong and CHEN Yongfu state key laboratory for Agrobiotechnology and Department of animal physiology,College of Biological Science,china Agricultural University,beijing 100094,china state key laboratory for Agrobiotechnology and Department of animal physiology College of Biological Science China Agricultural University Beijing 100094 China

Mouse early embryos undergo genome-wide demethylation and remethylation events during pre-implantation *** methylation reprogramming is thought to be associated with development *** immunofluorescence staining with an antibody against 5-methylcytosine (MeC),we examined the genome methylation patterns of mouse embryos cultured in *** results did not show the difference in staining patterns between development-blocked two-cell embryos that cultured in vitro and the two-cell embryos that were freshly collected from the donor *** in vitro-arrested morulae displayed a strong positive staining when compared to the morulae freshly collected from the donor *** the blastocyst stage,although most embryos showed the expected methylation patterns,with highly stained inner cell mass (ICM) and weekly stained trophectoderm (TE),a proportion of embryos were dimly stained in both ICM and *** results indicated that the methylation profile of the embryos could be changed by culturing in vitro when the embryos were in the transition from morulae to blastocyst.

关键词： methylation mouse embryo 5-methylcytosine antibody

Fine mapping quantitative trait loci affecting milk production traits on bovine chromosome 6 in a Chinese Holstein population

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Journal of Genetics and Genomics 2009年第11期36卷 653-660页

作者： Gui Mei 1,Cengceng Yin 1,Xiangdong Ding,Qin Zhang state key laboratory for Agrobiotechnology,key laboratory of animal Genetics and Breeding of the Ministry of Agriculture,College of animal Science and Technology,china Agricultural University,beijing 100193,china College of animal Science and Technology China Agricultural University State Key Laboratory for Agrobiotechnology Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture Beijing 100193

To fine map the previously detected quantitative trait loci （QTLs） affecting milk production traits on bovine chromosome 6 （BTA6）, 15 microsatellite markers situated within an interval of 14.3 cM spanning from BMS690 to BM4528 were selected and 918 daughters of 8 sires were genotyped. Two mapping approaches, haplotype sharing based LD mapping and single marker regression mapping, were used to analyze the data. Both approaches revealed a quantitative trait locus （QTL） with significant effects on milk yield, fat yield and protein yield located in the segment flanked by markers BMS483 and MNB209, which spans a genetic distance of 0.6 cM and a physical distance of 1.5 Mb. In addition, the single marker regression mapping also revealed a QTL affecting fat percentage and protein percentage at marker DIK2291. Our fine mapping work will facilitate the cloning of candidate genes underlying the QTLs for milk production traits.

关键词： fine mapping QTL Bos Taurus autosome 6 milk production traits

Effect of leptin on oocyte maturation and subsequent pregnancy rate of cloned embryos reconstructed by somatic cell nuclear transfer in pigs

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progress in Natural Science:Materials International 2008年第12期18卷 1583-1587页

作者： Hengxi Wei~a Qiuyan Li~(a,b) Jun Li~a Yan Li~b Yunping Dai~(a,b) Yufang Ma~a Kai Xue~a Ning Li~a a state key laboratory for Agrobiotechnology and Department of animal physiology,china Agricultural University,beijing 100094,china b Ji pulin Biotech Ltd.,beijing 100094,china state key laboratory for Agrobiotechnology and Department of animal physiology China Agricultural University Beijing 100094 China Ji pulin Biotech Ltd. Beijing 100094 China

Cloning pigs by somatic cell nuclear transfer (SCNT) has wide applications in basic research,human medicine and agricultural *** improve cloning efficiency,the effect of two basic maturation media,NCSU-23 and TCM199,was compared,and TCM199 was selected for the following experiments with *** systematically studied the effects of leptin supplementation on oocytes in vitro maturation (IVM),in vitro development of parthenogenetically activated (pA) and SCNT embryos and in vivo develop- ment of SCNT embryos after embryo transfer (ET).The results showed that supplementation of 100 or 200 ng/ml leptin into the mat- uration medium did not greatly affect nuclear maturation of oocytes,or cleavage rates of pA and SCNT (p>0.05).Blastocyst rates of pA and SCNT embryos were significantly improved when 100 or 200 ng/ml leptin was added to maturation medium,and the number of cells in pA blastocysts was also improved (pr of cells in blastocyst of SCNT was improved,when 100 ng/ml leptin was added (prthermore,supplementation of 100 or 200 ng/ml leptin to the IVM medium may improve pregnancy rate and the delivery rate in pig cloning.

关键词： Leptin Oocytes IVM SCNT pig

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%, prate of nuclear maturation of oocytes increased significantly when IVM duration time was extended from 36 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%, prmation (9.3% vs. 13.2%, prence was observed between NTEs reconstructed from oocytes matured under lower oxygen (7% O2) tension; NTEs derived from oocytes matured under higher oxygen tension (20% O2) in cleavage rate (70.6% vs. 67.1%, prate (11.8% vs. 12.3%, presults suggest that: (i) G418 selection does not have a significant effect on cleavage rate of NTEs expressing GFp. (ii) Nuclear maturation is greatly improved by prolonging IVM duration from 36 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.

reduction of drosopterin content caused by a 45-nt insertion in Henna pre-mrNA of Drosophila melanogaster

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Science china(Life Sciences) 2008年第8期51卷 702-710页

作者： WANG Qin, ZHAO ChunJiang, BAI LiHua, DENG XueMei & WU ChangXin state key laboratory of Agrobiotechnology & key laboratory of animal Genetics and Breeding of the Ministry of Agriculture, College of animal Science and Technology, china Agricultural University, beijing 100193, china state key laboratory of Agrobiotechnology & key laboratory of animal Genetics and Breeding of the Ministry of Agriculture College of Animal Science and Technology China Agricultural University Beijing China

phenylalanine hydroxylase is assumed to be a key enzyme in drosopterin metabolism, but direct in vivo evidence to support this hypothesis is still *** the present study, we found a new natural reces-sive purple eye mutant of Drosophila melanogaster, Hnbp, which was a 45-nt insertion mutant in the second exon of *** insertion resulted in a predicted protein with 15 additional amino acids as compared to the wild-type *** analysis of protein structure showed that the predicted mutant protein probably had two more β-sheets, which may cause instability of two α-helices near the catalytic centre of the enzyme in the Biopterin-Hydroxyl binding *** mutant showed eye color defect with decrease of mrNA level, as well as drosopterin content *** drosopterin defect could be fully rescued by expression of wild type Henna in the Hnbp background by GMr-GAL4 UAS-Henna/UAS-Henna:Hnbp/Hnbp transgenic *** taken together, it can be concluded that the mu-tation in Henna is responsible for drosopterin reduction in mutant Hnbp, which provides key in vivo evidence to support the hypothesis that Henna is involved in drosopterin synthesis.

关键词： Drosophila melanogaster Henna gene drosopterin phenylalanine hydroxylase Hnbp