以短串联重复序列（short tandem repeats, STR）和性别基因为基础的动物分型系统,不仅能提供重要的分子遗传标记信息,还能有效地解决动物个体识别和亲缘关系难题。为建立家鸽（Columba livia domestica）短串联重复序列与染色体螺旋蛋白基因（chromobox-helicase-DNA binding gene, CHD）复合扩增检测体系,并评估其法医学应用价值,本研究采用六色荧光标记复合扩增方法,选取了CliμD11、PG4、PG1、PG2、CliμT02、CliμD17、CliμD35、CliμT17、CliμD16、PIGN04、CliμD32、PIGN57、PIGN26、PG5、PG6、CliμD19、PIGN12、PIGN15、PIGN10和性别鉴定基因CHD,共计20个基因位点。结果显示,本研究建立的家鸽复合扩增体系的分型结果稳定、图谱清晰,241只家鸽的羽毛样本检测全部稳定可靠;检测灵敏度高达0.0625 ng;与其他物种对比扩增,表现出高度的种属特异性;对19个STR位点的等位基因在232只家鸽群体中的统计分析表明,本研究所选取的STR位点均可用于家鸽的个体识别和亲子鉴定。本研究最终成功建立的稳定性好、灵敏度高、种属特异性强的家鸽STR位点复合扩增检测体系,是目前已知位点数最多的家鸽检测体系,也是国内相关研究的首次报道,为家鸽的个体识别和亲子鉴定提供了分子鉴定基础。

Camalexin, a major phytoalexin in Arabidopsis thaliana, consists of an indole ring and a thiazole ring. The indole ring is produced from Trp, which is converted to indole-3-acetonitrile (IAN) by CYP79B2/CYP79B3 and CYP71A13. Conversion of Cys(IAN) to dihydrocamalexic acid and subsequently to camalexin is catalyzed by CYP71B15. Recent studies proposed that Cys derivative, not Cys itself, is the precursor of the thiazole ring that conjugates with IAN. The nature of the Cys derivative and how it conjugates to IAN and subsequently forms Cys(IAN) remain obscure. We found that protein accumulation of multiple glutathione S-transferases (GSTs), elevation of GST activity, and consumption of glutathione (GSH) coincided with camalexin production. GSTF6 overexpression increased and GSTF6-knockout reduced camalexin production. Arabidopsis GSTF6 expressed in yeast cells catalyzed GSH(IAN) formation. GSH(IAN), (IAN)CysGly, and γGluCys(IAN) were determined to be intermediates within the camalexin biosynthetic pathway. Inhibitor treatments and mutant analyses revealed the involvement of γ-glutamyl transpeptidases (GGTs) and phytochelatin synthase (PCS) in the catabolism of GSH(IAN). The expression of GSTF6, GGT1, GGT2, and PCS1 was coordinately upregulated during camalexin biosynthesis. These results suggest that GSH is the Cys derivative used during camalexin biosynthesis, that the conjugation of GSH with IAN is catalyzed by GSTF6, and that GGTs and PCS are involved in camalexin biosynthesis.

Summary Genome scans were conducted on an F 2 resource population derived from intercross of the White Plymouth Rock with the Silkies Fowl to detect QTL affecting chicken body composition traits. The population was genotyped with 129 microsatellite markers and phenotyped for 12 body composition traits on 238 F 2 individuals from 15 full‐sib families. In total, 21 genome‐wide QTL were found to be responsible for 11 traits, including two newly studied traits of proventriculus weight and shank girth. Three QTL were genome‐wide significant: at 499 c m on GGA1 (explained 3.6% of phenotypic variance, P < 0.01) and 51 c m on GGA5 (explained 3.3% of phenotypic variance, P < 0.05) for the shank & claw weight and 502 c m on GGA1 (explained 1.4% of phenotypic variance, P < 0.05) for wing weight. The QTL on GGA1 seemed to have pleiotropic effects, also affecting gizzard weight at 490 c m , shank girth at 489 c m and intestine length at 481 c m . It is suggested that further efforts be made to understand the possible pleiotropic effects of the QTL on GGA1 and that on GGA5 for two shank‐related traits.

High purity berry plasma membranes (PMs) of Vitis vinifera L. cv. Cabernet Sauvignon were isolated by two-phase partitioning of microsome fractions at different stages of berry ripening. PM proteins resolvable by the detergent cocktail of CHAPS and ASB-14 were separated by two-dimensional electrophoresis. A total of 119 protein spots from pre-véraison berry PMs on 2-D gels detected with silver staining were subjected to MALDI-TOF mass spectrometry analysis. Sixty-two spots were identified as putative PM proteins, with 1-6 predicted transmembrane helices, including true PM proteins such as ATP synthase, ABC transporters, and GTP-binding proteins reported in plants. They were then grouped into eight functional categories, mainly involved in transport, metabolism, signal transduction, and protein synthesis. Another 11 spots were identified as proteins of unknown function. The véraison and post-véraison samples stained 98 and 86 spots on the gels, respectively. During the berry ripening process, total PM protein content gradually decreased. Among all identified proteins, 12 showed significant differences in terms of their relative abundance. Increasing ubiquitin proteolysis and cytoskeleton proteins were observed from pre-véraison to post-véraison. Zeatin O-glucosyltransferase peaked at véraison, while ubiquitin-conjugating enzyme E2-21 was down-regulated at this stage. This proteome research provides the first information on PM protein characterization during the grape berry ripening process.

Insulin-like growth factor binding protein-2 is a member of the insulin-like growth factor families. Using a porcine RH panel, the gene was mapped on chromosome 15q22-23. Meanwhile, using polymerase chain reaction single strand conformation polymorphism, genotypic and allelic frequencies were analyzed in 17 pig breeds (total animals 570), together with a chi-square test of Hardy-Weinberg equilibrium. Also the association between haplotypes and production performance was analyzed in a Lantang x Landrace population family (n = 133, total 43 traits). At each locus we investigated, all the breeds showed different genotypic and allelic frequency distributions. In general, the Chinese native pig breeds carried a higher allele A frequency (over 50%) than the European pigs. For production performance, pigs with the CAG haplotype had higher fore-body and rear-body weight than those with the TGT and TAG haplotypes (P < 0.05). Also, pigs with the CAG haplotype had higher bone weight of the rear-body than those with the CAT haplotype (P < 0.05); pigs with the TGT and CAG haplotypes had higher forelimb and rearlimb weight than those with the CAT haplotype (P < 0.01 and P < 0.05, respectively); pigs with the TGG haplotype had higher leaf fat weight than those with the TGT and CAG haplotypes (P < 0.05); and pigs with the CAG haplotype had more stomach weight than those with the CAT and CGT haplotypes (P < 0.01); pigs with the TGT and CAG haplotypes had more ribs and longer body than those with the CGT-TGG, and CAT-TAG haplotypes (P < 0.05). These results suggest that IGFBP-2 is associated with production performance, but our population family was small. More studies with large samples are needed before the IGFBP-2 locus will be useful for a selection program.

We created a cDNA microarray representing approximately 3,500 pig genes for functional genomic studies. The array elements were selected from 6,494 cDNA clones identified in a large-scale expressed sequence tag (EST) project. These cDNA clones came from normalized and subtracted porcine adipose tissue cDNA libraries. Sequence similarity searches of the 3,426 ESTs represented on the array using BLASTN identified 2,790 (81.4%) as putative human orthologs, with the remainder consisting of "novel" genes or highly divergent orthologs. We used the gene microarray to profile transcripts expressed by adipose tissue of fatty Chinese Xiang pig (XP) and muscley Large White (LW). Microarray analysis of RNA extracted from adipose tissue of fatty XP and muscley LW identified 81 genes that were differently expressed two fold or more. Transcriptional differences of four of these genes, adipocyte fatty acid binding protein (aP2), stearyl-CoA desaturase (SCD), sterol regulatory element binding transcription factor 1 (SREBF1) and lipoprotein lipase (LPL) were confirmed using SYBR Green quantitative RT-PCR technology. Our results showed that high expression of SCD and SREBF1 may be one of the reasons that larger fat deposits are observed in the XP. In addition, our findings also illustrate the potential power of microarrays for understanding the molecular mechanisms of porcine development, disease resistance, nutrition, fertility and production traits.

采用中国泰和丝羽乌骨鸡和白洛克肉鸡的正反交组合构建了中国农业大学(CAU)资源家系,利用129个微卫星对资源群中的4个半同胞家系进行了大规模基因组扫描,构建了微卫星标记的连锁图谱(CAU遗传图谱),该图谱覆盖了23条常染色体(1-15,17-24,26和27)、1条性染色体(Z染色体)和2个连锁群(E26C13和E50C23),图谱总长为3307.5 cM;雄性与雌性的遗传图谱差异为3.51%.CAU遗传图谱的标记顺序与2000年发表的鸡的整合图谱的标记顺序一致,但是遗传长度有所不同.该图谱的构建为进一步的数量性状位点定位研究打下坚实基础.

A pig BAC library was constructed with genomic DNA from a male Erhualian pig. After partial digestion with Hind III or RamH I the 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 I and 8 with an average of 3.6. Positive superpools were obtained for 32 microsateltite 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 identification.

A genome scan to detect quantitative trait loci (QTL) affecting body weight in chickens was conducted on 238 F(2) chickens from a reciprocal cross of Silky Fowl and White Plymouth Rock using 125 microsatellite markers covering 23 autosomes and the Z chromosome. Two types of QTL were considered: static QTL (SQ) and developmental QTL (DQ). Static QTL affected the body weight from hatch to time t, and DQ affected the body weight from time t-1 to time t. Six SQ and nine DQ were detected. Of these QTL, four reached a genome-wide significance of 5% or better, with SQ1 and DQ1 being the most significant QTL. Static QTL1 was on chromosome 1 between GCT0006 and MCW0106 and explained 4.05-9.80% of the phenotypic variation in body weights from 3 to 12 weeks of age. At 9, 10 and 11 weeks, the genome-wide significance thresholds of SQ1 were <1%. Developmental QTL1 was located on chromosome 1 between MCW0168 and GCT0006, and explained 2.75% of the phenotypic variation for body weight from week 7 to 8 with a genome-wide significance level <1%. The results suggest that body weight from hatch to time t and developmental growth from time t-1 to time t may involve two different sets of genes or gene actions.

In order to study duck microsatellites, we constructed a library enriched for (CA)n, (CAG)n, (GCC)n and (TTTC)n. A total of 35 pairs of primers from these microsatellites were developed and used to detect polymorphisms in 31 unrelated Peking ducks. Twenty-eight loci were polymorphic and seven loci were monomorphic. A total of 117 alleles were observed from these polymorphic microsatellite markers, which ranged from 2 to 14 with an average of 4.18 per locus. The frequencies of the 117 alleles ranged from 0.02 to 0.98. The highest heterozygosity (0.97) was observed at the CAUD019 microsatellite locus and the lowest heterozygosity (0.04) at the CAUD008 locus, and 11 loci had heterozygosities greater than 0.50 (46.43%). The polymorphism information content (PIC) of 28 loci ranged from 0.04 to 0.88 with an average of 0.42. All the above markers were used to screen the polymorphism in other bird species. Two markers produced specific monomorphic products with the chicken DNA. Fourteen markers generated specific fragments with the goose DNA: 5 were polymorphic and 9 were monomorphic. But no specific product was detected with the peacock DNA. Based on sequence comparisons of the flanking sequence and repeat, we conclude that 2 chicken loci and 14 goose loci were true homologous loci of the duck loci. The microsatellite markers identified and characterized in the present study will contribute to the genetic map, quantitative traits mapping, and phylogenetic analysis in the duck and goose.

A 273 base pair (bp) fragment of the SLA-DQB gene including parts of intron 1 and exon 2 has been investigated using PCR-RFLP in 38 indigenous Chinese pig breeds, two Chinese wild boars, and three foreign pig breeds. The restriction enzyme RsaI revealed three polymorphic sites in the 273 bp fragment for the pig breeds studied. In total, four alleles resulting in 10 genotypes were found. Twenty pig breeds are not in Hardy–Weinberg equilibrium at this locus. The allele frequency of a chi-square test showed that there is significant difference (P < 0.05) among six Chinese pig groups, and an even greater significant difference (P < 0.01) was found between Chinese and European pig breeds.

For the first time, a 16 kb fragment of the porcine Ob gene, namely intron1, exon1, 5' region of Ob gene, was restrictively analyzed and sequenced by the primers designed in the portion of the swine Ob sequence. The first small 38 bp untranslated exon1 is located 11.1 kb upstream of the initiator ATG codon, and two novel microsatellites SW200 and SW160 are found in intron1. Promoter analysis of several putative binding sites revealed that this 300 bp promoter located at -1 to -300, including C/EBP and two Sp1, may be as effective as the longer promoter in directing leptin transcription. To examine microsatellites association with important economic traits, we statistically analyzed genotypes and alleles of the two microsatellites. Statistical analysis carried out by SAS 8.2 revealed significant positive correlation between the two microsatellites genotypes and the litter size in first parity of Erhualian.

Animal GeneticsVolume 36, Issue 3 p. 284-284 Linkage mapping of the SCN8A gene to chicken linkage group E22C19W28 Y. Gao, Y. Gao State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this authorX. X. Hu, X. X. Hu State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this authorX. M. Deng, X. M. Deng State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this authorJ. D. Feng, J. D. Feng State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this authorN. Li, N. Li State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this author Y. Gao, Y. Gao State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this authorX. X. Hu, X. X. Hu State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this authorX. M. Deng, X. M. Deng State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this authorJ. D. Feng, J. D. Feng State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this authorN. Li, N. Li State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, ChinaSearch for more papers by this author First published: 02 June 2005 https://doi.org/10.1111/j.1365-2052.2005.01302.xCitations: 4 Ning Li ([email protected]) Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. References 1 Plummer N. W. et al. (1998) Genomics 54, 287–96. 2 Kearney J. A. et al. (2002) Hum Mol Genet 11, 2765–75. 3 Meisler M. H. et al. (2004) Genetica 122, 37–45. 4 Hubbard T. et al. (2002) Nucleic Acids Res 30, 38–41. 5 Deng X. M. et al. (2000). Yi Chuan Xue Bao 28, 801–7. 6 Green P. et al. (1990) Documentation for CRI-MAP, Version 2.4. Washington School of Medicine, St Louis, MO. 7 Groenen M. A. M. et al. (2000) Genome Res 10, 137–47. 8 Schmid M. et al. (2000) Cytogenet Cell Genet 90, 169–218. 9 Bitgood J. J. et al. (1993) Genetic Maps, 6th edn. (Ed. by S. O'Brein ), pp. 4332–42. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, NY. 10 Ruyter-Spira C. P. et al. (1997) Poult Sci 76, 386–91. Citing Literature Volume36, Issue3June 2005Pages 284-284 ReferencesRelatedInformation

Avian infectious bronchitis virus (AIBV) is classified as a member of the genus coronavirus in the family coronaviridae. The enveloped virus has a positive-sense, single-stranded RNA genome of approximately 28 kilo-bases, which has a 5′ cap structure and 3′ polyadenylation tract. The complete genome sequence of infectious bronchitis virus (IBV), Beijing isolate , was determined by cloning sequencing and primer walking. The whole genome is 27733 nucleotides in length, has ten open reading frames: 5′ - orf1a-orf1ab-s-3a-3b-e-m-6a-6b-n-3′ . Alignments of the genome sequence of IBV Beijing isolate with those of two AIBV strains and one SARS coronavirus were performed respectively. The genome sequence of IBV Beijing isolate compared with that of the IBV strain LX4 (uncompleted, 19440 bp in size) was 91.2% similarity. However, the full-length genome sequence of IBV Beijing isolate was 85.2% identity to that of IBV Strain Beaudette , and was only 50.8% homology to that of SARS coronavirus. The results showed that the genome of IBV has remarkable variation. And IBV Beijing isolate is not closely related to SARS coronavirus. Phylogenetic analyses based on the whole genome sequence, S protein, M protein and N protein, also showed that AIBV Beijing isolate is lone virus in group III and is distant from SARS coronavirus. In conclusion, this study will contribute to the studies of diagnosis and diseases control on IBV in China.

鸟类传染性支气管炎病毒(AIBV)属于冠病毒科, 冠状病毒属, 是单股线状、正链RNA病毒, 基因组全长近28 kb, 具有3′多聚A尾和5′帽子结构的特征. 采用PCR产物克隆测序和引物步移(primer walking)直接测序结合的方法, 完成了IBV北京分离株的全基因组序列的测定. 该毒株基因组序列全长为27733 bp, 生物信息学分析表明, 它具有10个明显的可读框(ORF); 通过基因定位后其基因排序为: 5′-orf1a-orf1ab-s-3a-3b-e-m-6a-6b-n-3′. 对IBV北京分离株的全基因组序列, 与报道的IBV及造成人类严重急性呼吸综合征(SARS)的病毒(SARS-CoV)进行了比较分析. 结果表明, AIBV是一种变异较大的病毒, 其全基因组序列与国外公布的AIBV全基因组序列相似性仅有85.2%, 与国内报道的不完整AIBV序列比较, 相似性也只有91.2%; 而与SARS病毒基因组全序列比较, 相似性仅为50.8%, 说明它与SARS-CoV关系不大. 同时, 还使用clustalw 1.81 和 Treeview软件构建了冠状病毒的全序列、S蛋白、M蛋白和N蛋白的系统发生树, 结果表明, 鸡IBV北京分离株是第3组病毒的惟一成员, 它与SARS-CoV存在很大的遗传距离. 这项研究将对我国鸡传染性支气管炎病原的鉴定和疾病的控制起到重要的作用.

RNA病毒在复制时有易出错的倾向,因此SARS病毒在传染或传代过程中易发生突变而产生不同的毒株,这种机制也利于病毒逃脱宿主的免疫系统而生存下来.许多研究也表明,不同的SARS病毒分离株的全基因组序列存在不同程度的差异,这些差异的研究无疑对SARS病毒疫苗的研制具有重要的指导意义.同时,对SARS病毒在传代过程中的遗传稳定性及核酸特性的分析,是SARS疫苗研制不可或缺的环节.采用PCR产物直接测序的方法,对2株用Vero细胞培养经过多次传代的SARS病毒进行了全基因组序列的测定,通过比较分析发现该病毒在传代过程中具有高遗传稳定性,其中检定参比株(Sino1株)2和11代全基因组序列比较有4个碱基的变化,而候选疫苗株(Sino3株)3与10代的基因组全序列仅有1个碱基的差异.SARS病毒在Vero细胞上传代的遗传稳定性表明,以此制备的灭活病毒疫苗是稳定的.

Small Tail Han Sheep has significant characteristics of high prolificacy and non-seasonal ovulatory activity and is an excellent local sheep breed in P. R. China. Recently a novel member of the transforming growth factor β (TGFβ) superfamily termed bone morphogenetic protein 15 (BMP15) was shown to be specifically expressed in oocytes and to be essential for female fertility. Therefore, BMP15 is a candidate gene for reproductive performance of Small Tail Han Sheep. The whole genomic nucleotide sequence of BMP15 gene in Small Tail Han Sheep was searched for polymorphisms by PCR-SSCP and direct sequencing, and only one polymorphism was found. The polymorphism was a result of a 3 base pair deletion, which eliminated a single Leu codon (CTT). The allelic frequencies for A (without deletion) and B (with a codon deletion) are 0.73 and 0.27 respectively. The effects of BMP15 genotype on litter size were evaluated using the least squares model. This indicated that there was a significant association between litter size of Small Tail Han Sheep and a deletion in BMP15 gene (p=0.02<0.05). Small Tail Han Sheep ewes with AA and AB genotype produce on average 0.5 and 0.3 more lambs per litter than those ewes with BB genotype.

SARS coronavirus is an RNA virus whose replication is error-prone, which provides possibility for escape of host defenses, and even leads to evolution of new viral strains during the passage or the transmission. Lots of variations have been detected among different SARS-CoV strains. And a study on these variations is helpful for development of efficient vaccine. Moreover, the test of nucleic acid characterization and genetic stability of SARS-CoV is important in the research of inactivated vaccine. The whole genome sequences of two SARS coronavirus strains after passage in Vero cell culture were determined and were compared with those of early passages, respectively. Results showed that both SARS coronavirus strains have high genetic stability, although nearly 10 generations were passed. Four nucleotide variations were observed between the second passage and the 11th passage of Sino1 strain for identification of SARS inactivated vaccine. Moreover, only one nucleotide was different between the third passage and the 10th passage of Sino3 strain for SARS inactivated vaccine. Therefore, this study suggested it was possible to develop inactivated vaccine against SARS-CoV in the future.