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Zhang Z.-J.,Anhui Agricultural University | Zhang Z.-J.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | Ling Y.-H.,Anhui Agricultural University | Ling Y.-H.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | And 12 more authors.
Asian Journal of Animal and Veterinary Advances | Year: 2013

Myostatin (MSTN) was a major gene of skeletal muscle growth regulation and a member of the transforming growth factor-β (TGF-β) super family. It acts as a negative regulator of skeletal muscle growth. To knock out goat MSTN gene of nuclear donor cells correctly, a gene targeting vector should be constructed. The homologous arm of goat MSTN was amplified by PCR and restriction enzyme digested. Intermediate vector pMD18-SA and pMD18-LA were constructed by conventional molecular cloning method. The final target vector pLOXP-MSTN was constructed by step-by-step cloning. The replacement type knock-out vector was transfected into goat fetal fibroblast by electrotransfection. The homologous arm sequences of goat MSTN including the 4.6 kb homologous long arm (LA) and the 1.9 kb homologous Short Arm (SA) were successfully amplified using LA-PCR technology. The LA and SA were inserted into vector pLOXP, generating MSTN gene targeting vector pLOXP-MSTN which contained neo and tk positive-negative-selection markers. The pLOXP-MSTN was linearized and electroporated into the nuclear donor cells. After G418 and GANC selection, fifty-eight drug resistant cell clones were screened and from which totally 3 positive clones were confirmed by PCR and DNA sequencing. The constructed gene targeting vector could efficiently target of MSTN locus and the gene knockout cell clones will be used to produce MSTN knockout goat by means of somatic cell nuclear transplantation. © 2013 Academic Journals Inc.


Ling Y.H.,Anhui Agricultural University | Ling Y.H.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | Quan Q.,Anhui Agricultural University | Xiang H.,Anhui Agricultural University | And 8 more authors.
Genetics and Molecular Research | Year: 2015

Although RNA-Seq is an effective method for identifying and exploring novel functional genes in mammals, it has rarely been applied to study fertility-related genes in the goat. In this study, RNA-Seq was used to screen the estrus ovaries of uniparous and multiparous Anhui white goats (AWGs). In total, 15,890 genes were identified and 2201 of these were found to be differentially expressed between the genetic libraries from uniparous and multiparous goats. Compared to the uniparous library, 1583 genes were up-regulated and 618 genes were down-regulated in the multiparous library. The FER1L4 gene showed the level of highest up-regulation in the multiparous library, while SRD5A2 expression showed the greatest down-regulation. In order to determine the functions of FER1L4 and SRD5A2 in goats, the expression profiles of the two genes in different tissues from AWGs and Boer goats at diestrus were analyzed by quantitative PCR. FER1L4 and SRD5A2 showed tissue specific expression patterns and were highly expressed in ovaries from both AWGs and Boer goats. FER1L4 was more highly expressed in ovaries from multiparous than uniparous AWGs. In contrast, SRD5A2 was expressed at a lower level in multiparous AWGs. These results indicated that FER1L4 and SRD5A2 may be associated with the high fecundity of AWGs. © FUNPEC-RP.


Jin H.,Anhui Agricultural University | Jin H.,Hunan Agricultural University | Chen H.-Q.,Anhui Agricultural University | Chen H.-Q.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | And 9 more authors.
Asian Journal of Animal and Veterinary Advances | Year: 2012

Adenosine Monophosphate activated Protein Kinase (AMPK) plays a key role in cellular metabolism and regulating energy and its y3 subunit was encoded by the PRKAG3 gene which was regarded as the candidate gene for carcass quality trait in domestic animals and fowls. To understand PRKAG3 gene variation and its association with carcass quality of goat breeds, the polymorphism of goat PRKAG3 gene and its distribution patterns in different goat breeds were investigated. Five goat breeds selected were Anhui white goat, Matou, Boer, Xiangdong and Sanen dairy goat. DNA samples of all goat breeds were extracted from goat ear tissue. The polymorphisms in the 5' regulatory region and exon 13 of PRKAG3 gene were examined using Apal, Hpyl88I, Acil and Clal enzyme digestion, respectively and their distribution patterns were analyzed in different goat breeds. The results showed that two mutation loci in the 5' regulatory region, C-525A and C-225T, located at -525 and -225 bp upstream of the start codon of the PRKAG3 gene and two mutation loci in the exon 13, T90C and C102T, located at 90 bp and 102 bp of the exon 13. The 525CC genotype frequencies of five goat breeds listed above were 0.8000, 0.9681, 0.6042, 0.7557 and 0.8889, respectively, while 225CC genotype frequencies corresponding to 0.7391, 0.9750, 0.8684, 0.9785 and 0.9344. The mutations at T90C and C102T didn't cause the substitution of corresponding amino acids in the AMPK protein. The muscle fat grade and carcass fat grade are lower in goat breeds with higher 525CC225CC genotype frequency while higher in the breeds with higher 525CA225CT genotype frequency (p<0.05). The results suggest that the lipoidosis ability of goat breeds may be associated with C-525A and C-225T loci of PRKAG3 gene. © 2012 Academic Journals Inc.


Ling Y.H.,Anhui Agricultural University | Ling Y.H.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | Xiang H.,Anhui Agricultural University | Xiang H.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | And 12 more authors.
Genetics and Molecular Research | Year: 2014

The desmoglein 4 (DSG4) gene is a potential candidate in the search for genes that may affect wool traits, because of its function. This study aimed to screen for polymorphisms in partial exon 16 and 3'UTR of the sheep desmoglein 4 DSG4 gene, and to test its possible association with wool length and crimp associated with fur. Overall, 326 sheep were scanned via single-strand conformational polymorphism assay, through three pairs of primers. The breeds included Tan, Han, and TanxHan from China, Polled Dorset from Australia, and Suffolk from Britain genotypes AA, BB, and AB for primer2 and genotypes DD, EE, and DE for primer3 were detected in native breeds. Six SNPs and 3-bp insertion/deletions were found in exon 16, of which 4 lead to amino acid substitutions. In addition, 1 SNP was found in 3'UTR. The DSG4 genotype was found to be strongly associated with all wool traits that were considered in this study (P < 0.01). Sheep with the genotype MM had a higher least square mean compared to sheep with the genotype WW or WM with respect to birth scapular wool length (P < 0.01), crimp number of birth scapular wool crimp (P < 0.01), crimp number of weaning scapular wool crimp (P < 0.01), and crimp number of weaning rump wool crimp (P < 0.01, P < 0.05). In conclusion, our study is the first to demonstrate that the DSG4 gene may be a candidate, or major gene, which influences important wool traits. © FUNPEC-RP.


Ling Y.H.,Anhui Agricultural University | Ling Y.H.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | Ling Y.H.,Chinese Academy of Agricultural Sciences | Zhang X.D.,Anhui Agricultural University | And 15 more authors.
Asian-Australasian Journal of Animal Sciences | Year: 2012

To investigate the genetic diversity of seven Chinese indigenous meat goat breeds (Tibet goat, Guizhou white goat, Shannan white goat, Yichang white goat, Matou goat, Changjiangsanjiaozhou white goat and Anhui white goat), explain their genetic relationship and assess their integrity and degree of admixture, 302 individuals from these breeds and 42 Boer goats introduced from Africa as reference samples were genotyped for 11 microsatellite markers. Results indicated that the genetic diversity of Chinese indigenous meat goats was rich. The mean heterozygosity and the mean allelic richness (AR) for the 8 goat breeds varied from 0.697 to 0.738 and 6.21 to 7.35, respectively. Structure analysis showed that Tibet goat breed was genetically distinct and was the first to separate and the other Chinese goats were then divided into two sub-clusters: Shannan white goat and Yichang white goat in one cluster; and Guizhou white goat, Matou goat, Changjiangsanjiaozhou white goat and Anhui white goat in the other cluster. This grouping pattern was further supported by clustering analysis and Principal component analysis. These results may provide a scientific basis for the characteristization, conservation and utilization of Chinese meat goats.


Zhang Z.J.,Anhui Agricultural University | Zhang Z.J.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | Ling Y.H.,Anhui Agricultural University | Ling Y.H.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | And 12 more authors.
Genetics and Molecular Research | Year: 2013

Mutations in the myostatin (MSTN) gene can inactivate its expression and result in a non-functional protein, which leads to dramatic muscularity and a "double-muscling" phenomenon in many species. Using gene sequencing and polymerase chain reaction-single-strand conformation polymorphism methods, polymorphisms of the MSTN gene were investigated as a candidate marker for growth in 288 goats. The results showed 2 novel single nucleotide polymorphisms: DQ167575 g.197G>A and 345A>T. Three potential genotypes (AA, AB, and BB) of substitution 197G>A in the 5'-untranslated region were detected in the 2 breeds. The polymorphism (CC and CD) of substitution 345A>T in exon I was segregated. The genetic diversity analysis revealed that Boer goat and Anhui white goat possessed intermediate genetic diversity in the P1 and P3 loci. Significant associations between the genotypes of the P3 locus and body weight, body length, and body height were observed in Boer goat and Anhui white goat (P < 0.05). It could be inferred that the MSTN gene may be a major gene or linked to the major gene affecting the goat growth traits. The polymorphic site could be a molecular marker-assisted selection program for body weight. © FUNPEC-RP.


Ling I.Y.H.,Anhui Agricultural University | Ling I.Y.H.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | Ding J.P.,Anhui Agricultural University | Ding J.P.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | And 10 more authors.
Research Journal of Biotechnology | Year: 2015

Many miRNAs have been reported for their important roles in developmental processes in various animals but there is limited information about goat miRNAs. In this study, the temporal and spatial expression patterns of miR-206 in skeletal muscle (longissimus dorsi) from Anhui white goat and Boer goat were assessed using Real time quantitative PCR (qPCR) method. In addition, the association between miR-206 expression and the muscle development was analyzed. The expression of miR-206 has tissue and time specificity. The expression level in longissimus dorsi muscle from 360 days goats was the highest while the expression level in neonate was the lowest. In 180 days goats, miR-206 was abundantly expressed in muscle and heart, moderately in liver and kidney and weakly in other collected tissues. The results indicated that miR-206 may be involved in myofibre proliferation and differentiation and further contribute to skeletal muscle development and phenotype with different genotypes. The present results provided the theory base for study of miR-206 function.


Zhang Z.,Anhui Agricultural University | Zhang Z.,Local Animal Genetic Resources Conservation and Biobreeding Laboratory of Anhui Province | Chen J.,Anhui Agricultural University | Huang Y.,Anhui Agricultural University | And 4 more authors.
Nongye Gongcheng Xuebao/Transactions of the Chinese Society of Agricultural Engineering | Year: 2013

In order to ascertain the environmental condition of goat shed and its existing problems in South China, this study had been conducted for two years to detect and analyze the air environmental indicators in double-sloped airtight goat houses with windows and slatted floor, which was the most commonly seen type of goat house in Southern China, in summer in Jianghuai Region. The results showed that: (1) daily mean temperature, relative humidity, airflow velocity, illuminance, noise and concentration of nitrogen and carbon dioxide in such goat houses in the hottest month in summer were 30.5°C, 70.2%, 0.16 m/s, 99.45 lx, 49.82 dB, 1.98 mg/m3, 0.060% respectively. All indicators met the required standards of environmental hygiene except for the daily mean temperature. (2)Measurement in different sampling points of goat houses showed that characteristics of space distribution of each above-mentioned parameters were not very apparent. (3)As outside environment changed, daily mean temperatures and relative humidity in the sampling points also changed apparently. The shift was relatively steady in January, while temperatures in rainy days dropped fairly significantly (6.9°C). Average temperature difference between days and nights during the trial period was 2.3°C. Average indoor and outdoor temperature difference in the goat houses was 1.8°C. (4) Daily and monthly concentration of nitrogen and carbon dioxide in the sampling sites shifted slightly. (5)There was a significant negative correlation between temperature and relative humidity of the air in the sampling sites (r=-0.9818, P<0.01), while a significant positive correlation was detected between the concentration of nitrogen and the temperature and relative humidity of the air (r=0.8702, P<0.05). It is recognized based on the comprehensive evaluation that microclimatic environment of double-sloped airtight goat house with windows and slatted floor is basically in accordance with biological characteristics of environmental hygiene and goats. Thus, this type of goat house is suitable to be promoted and applied in Jianghuai Region, and it is only necessary to take certain measures to lower the temperature in the houses in hot summer weather.

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