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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%, p0.05). 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%, p<0.05); blastocyst formation (9.3% vs. 13.2%, p<0.05); (iii) no significant difference 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%, p<0.05); blastocyst rate (11.8% vs. 12.3%, p<0.05). These results 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.

QTL alleles on chromosome 7 from fatty Meishan pigs reduce fat deposition

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Science China(Life Sciences) 2003年第1期46卷 10-17页

作者：岳根华 Petra Beeckmann Gerhard Moser Elisabeth Müller Hans Bartenschlager1 Stanislav Cepica Jaroslav Schrffel Antonin Stratil Hermann Geldermann Department of Animal Breeding and Biotechnology University of Hohenheim Institute of Animal Physiology and Genetics Academy of Sciences of the Czech Republic

For detecting QTL in the whole swine genome,1068 pigs from three F2 populations con-structed by crossing European Wild boar and Pietrain (W譖),Meishan and Pietrain (M譖),and Wild Boar and Meishan (W譓) were genotyped for genetic markers evenly spaced at approximately 20 cM inter-vals. A QTL analysis was performed using a least-squares method. Here the results of the QTL analysis on the porcine chromosome 7 are presented. QTL for carcass composition (*** weight,carcass length,backfat depth,abdominal fat and bacon meat) were mapped in the chromosomal region CYPA/CYPD-TNFB-S0102 in M譖 and W譓,but not in W譖. The QTL was additive. Surprisingly, in contrast to the parental phenotype, the QTL alleles from fatty Meishan were associated with thinner backfat than Pietrain and Wild Boar alleles,suggesting that the genome of the fatty Meishan pig contains genes which can reduce fat content of carcass substantially.

关键词： genomics, pig, SSC7, QTL, backfat.

Cloning and analysis of differentially expressed ESTs in swine muscle tissue

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Science China(Life Sciences) 2006年第4期49卷 342-348页

作者： LI Chongsheng CHEN Yaosheng WANG Chong LI Jiaqi College of Animal Science South China Agricultural University Guangzhou 510642 China College of Animal Science South China Agricultural University Guangzhou 510642 China College of Animal Science South China Agricultural University Guangzhou 510642 China College of Animal Science South China Agricultural University Guangzhou 510642 China

The obvious difference in muscle growth; meat quality traits exists between Chinese indigenous pig; exotic pigs. In order to study the reason of these phenotypic differences; search the potential gene related to growth; meat quality traits, silver-stained mRNA differential display technique was used to detect the difference with mRNA of loin-eye muscle tissue from maturity pigs of Lantang in Guangdong Province; Large Yorkshire. One of the newly discovered expressed sequence tag (ESTsp3) was analyzed by using bioinformatic technique. The results showed: (1) nearly 2000 cDNA fragments were detected with 30 primer pairs,; 6 differentially expressed ESTs in the Ioin-eye muscle tissues from the two breeds were isolated; obtained. The differential fragments were cloned; sequenced. The all sequences were recorded in the GenBank. (2) The 786 bp fragment of ESTsp3 was obtained with in silico elongation system, the ORF analysis revealed that it existed as an 83 aa complete open reading frame,; the elongation sequences were verified by RT-PCR. The analysis of in silico expression profile showed that ESTsp3 is expressed in various growth stages; in most tissues; organs, such as soft tissue, skin, skeletal muscle; kidney, but with variant expression quantity.

关键词： pig, muscle, mRNA differential display, in silico expression profile.

Polymorphisms in the 5 ′ regulatory region of my-ostatin gene are associated with early growth traits in Yorkshire pigs

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Science China(Life Sciences) 2007年第5期50卷 642-647页

作者： YU LingZhi1, TANG Hui1, WANG JiYing2, WU Ying2, ZOU LiLi1, JIANG YunLiang1, WU ChangXin3 & LI Ning4 1 College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Taian 271018, China 2 Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan 250100, China 3 College of Animal Science and Technology, China Agricultural University, Beijing 100094, China 4 State Key Laboratory for Agro-biotechnology, China Agricultural University, Beijing 100094, China. College of Animal Science and Veterinary Medicine Shandong Agricultural University Taian 271018 China Institute of Animal Science and Veterinary Medicine Shandong Academy of Agricultural Sciences Jinan 250100 China College of Animal Science and Technology China Agricultural University Beijing 100094 China State Key Laboratory for Agro-biotechnology China Agricultural University Beijing 100094 China

Myostatin is a negative regulator of skeletal muscle *** present studycloned the 5' regulatory region of porcine myostatin gene,screened its polymorphisms and analyzedtheir associations with early growth traits in Yorkshire *** results indicated that a fragmentlength polymorphism and a polymorphism concerning two nucleotide changes exist in the 5' regulatoryregion of porcine myostatin *** sites 435 and 447,allele A and allele B have the haplotypes ofA-G and G-A,*** allelic frequency of B is 0.475 in Yorkshire *** homozygous BBgenotype was detected in 9 Laiwu Black *** B was found to have positive effect on bodyweight on day 21 (BW21) (P<0.01),body weight on day 28 (BW28) (P<0.05),body weight on day 70 (BW70)(P<0.05),average daily gain from birth to 21 d (ADG1) (P<0.01),average daily gain from birth to 28 d(ADG2) (P<0.05) and average daily gain from 21 d to 70 d (ADG3) (P<0.01),*** additiveeffect of allele B on BW21,BW28,BW70,ADG1,ADG2 and ADG3 was 0.596±0.205 kg (P=0.0041),0.498±0.200kg (P=0.0136) 1.409±0.551 kg (P=0.0112),28.39±9.74 g P=0.0041),17.78±7.15 g (P=0.0136) and37.00±16.92 g (P=0.0304),respectively,whereas its effect on average daily gain from 28 d to 70 d(ADG4) was not significant (P>0.1),although BB individuals are superior in average daily gain to AAand AB.

关键词： pig, myostatin, 5' regulatory region, polymorphism, early growth trait

Gene expression profiling in porcine mammary gland during lactation and identification of breed- and developmental-stage-specific genes

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Science China(Life Sciences) 2006年第1期49卷 26-36页

作者： SU Zhixi1, DONG Xinjiao2, ZHANG Bing1, ZENG Yanwu1, FU Yan1, YU Jun1,3 & HU Songnian1,3 1. James D. Watson Institute of Genome Sciences, Zhejiang University, Hangzhou 310008, China 2. School of Life and Environmental Sciences, Wenzhou Normal College, Wenzhou 325027, China 3. Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 101300, China James D. Watson Institute of Genome Sciences Zhejiang University Hangzhou 310008 China School of Life and Environmental Sciences Wenzhou Normal College Wenzhou 325027 China James D. Watson Institute of Genome Sciences Zhejiang University Hangzhou 310008 China Beijing Institute of Genomics Chinese Academy of Sciences Beijing 101300 China

A total of 28941 ESTs were sequenced from five 5′-directed non-normalized cDNA libraries, which were assembled into 2212 contigs; 5642 singlets using CAP3. These sequences were annotated; clustered into 6857 unique genes, 2072 of which having no functional annotations were considered as novel genes. These genes were further classified into Gene Ontology categories. By comparing the expression profiles, we identified some breed-and developmental-stage-specific gene groups. These genes may be relative to reproductive performance or play important roles in milk synthesis, secretion; mammary involution. The unknown EST sequences; expression profiles at different developmental stages; breeds are very important resources for further research.

关键词： expressed sequence tags, expression profile, domestic pig, mammary gland.