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Articles by Wenjin Zhu
Total Records ( 8 ) for Wenjin Zhu
  Zhengzhu Liu , Yuanfang Gong , Wenjin Zhu , Lingxin Duan , Muxiang Ge , Baochang Shi , Minshan Feng and He Gao
  In order to explore the sequence structure of fox Agouti gene and it’s mechanism to regulate the pelage color’s dividing. In this study, the major part of intron 2 sequence (1038 bp) of Agouti gene from the silver fox were obtained by PCR amplification and direct sequencing. This sequence was aligned with red fox, giant panda, horse, pig, goat, cattle, sheep, domestic cat and rabbit and the sequence similarities were 100, 85.82, 75.75, 73.31, 66.22, 65.98, 65.77, 60.45 and 58.82%, respectively. The result of the homology analysis showed that the genetic relationship between silver fox and red fox was the highest, which was consistent with that they belong to Vulpes of the Canidae animal in traditional classification. Based on the sequence of Agouti gene intron 2, the phylogenetic tree was constructed for silver fox and the other 9 species using Clustalx (1.83) software. The cluster result of phylogenetic tree of all species was basically consistent with the taxonomy of NCBI and was similar to the physiological characteristics of the species and their traditional classification. The above results provide the important biological information for researching the mechanism of the formation mechanism of the coat color and artificially improving the coat color quality of fox and so on.
  Zhengzhu Liu , Yuanfang Gong , Lingxin Duan , Minshan Feng , Wenjin Zhu , Muxiang Ge , Xierong Liu , Junsheng Zhao and Xinling Guo
  To explore the sequence structure of fox TYR gene and it’s mechanism to regulate the pelage color’s dividing. In this study, the partial DNA sequence (1370 bp) of silver fox TYR gene was obtained by PCR amplification and direct sequencing. This sequence contained the intron 1 of 368 bp, the complete exon 2 of 217 bp and the intron 2 of 785 bp and GC content was 40.5%. Based on the sequences of TYR gene exon 2, the homological analysis and the construction of un-rooted phylogenetic tree were carried out among silver fox and the other 16 species by using DNAMAN software and Clustalx (1.83) software, respectively. The homological analysis showed that silver fox had the closest genetic relationship with dog and the closer genetic relationship with domestic cat and domestic ferret. The cluster result of un-rooted phylogenetic tree was similar to that of the homological analysis. The length of TYR gene exon 2 were 217 bp for all analyzed species, which indicated that TYR gene exon 2 were highly conservative in phylogeny. The above results can provide the important biological information for further researching the development and regulation mechanism of fox coat color which result from TYR gene.
  Wenjin Zhu , Yongmei Su and Jianhua Wu
  The PCR primers were designed online according to result of gene homology comparison. Donkey GH gene DNA and cDNA sequence were cloned from liver and blood by RT-PCR and PCR, compared with GH gene sequences of different species by bioinformatics. The donkey GH gene sequence was 1928 bp including the 706 bp cDNA sequence with the complete CDS. By comparison for DNA and cDNA sequences, it was found that the GH gene sequence included 5 exons and 4 introns, encoding 216aa including signal protein of 26aa and matured protein of 190aa. Based on the analysis of the similarity of GH genes in different species on the level of cDNA, DNA and the deduced amino acid there was the most homology to the horse. The GH gene of donkey was conservative in the process of evolution and its promotor was not specific TATA box but was CATA box. The mutation of C→G in 1267 may affect the growth and development of donkeys and horses. All researches made an essential foundation for GH regulation of gene expression, evolution, polymorphism analysis in the future.
  Wenjin Zhu , Yongmei Su , Yanhua Ma , Jing Ni , Xuemin Guan and Jianhua Wu
  To reveal the genetical diversity of different donkey breeds at molecular level and provid some bases for the breed resource evaluation and utilization of donkeys in China, polymorphism of the second intron of the GH gene was analysised by PCR-SSCP in the seven donkey breeds (LX, GZ, XJ, GL, HB, DZ, JN). The results showed three haplotypes with the percentage of 1.7% in 174 samples were obtained, the haplotype diversity was high as 0.678 and 0.542 for the Linxian and Huaibei donkey and low as 0.077 for Xingiiang donkey with that of 0.409 and 0.462 for Guangling donkey and Guanzhong donkey, respectively with that of 0.355 and 0.304 for Dezhou donkeys and Jinnan donkeys, respectively. The amplified fragments of A and B haplotypes were cloned and sequenced. The result showed the fragments of B haplotypes had one substitution mutation at 735 site (G→C), the fragments of A haplotypes had one substitution mutation at 869 site (G→T). The results confirmed that there were polymorphisms in the second intron (214 bp) of donkey GH gene for the first time.
  Yongmei Su , Wenjin Zhu and Jianhua Wu
  This experiment was conducted to study the polymorphism on intron 3 of GH gene in donkey in China. The polymorphism was analysised by PCR-SSCP in the 10 donkey breeds (JN, GL, GZ, DZ, JM, BY, QY, LZ, TH and XJ) by PCR-SSCP. One SNPs was found in the sequence of GH gene through the sequencing analysis of the two homozygous genes AA and BB, the mutation at 100th site (G→T) of the third intron of donkey GH gene was showed. Three genotypes AA, AB and BB were detected in 10 donkey breeds and the allele A was the predominant allele. All the breeds were in Hardy-Weinberg equilibrium at this polymorphic site (p>0.05). The results confirmed that there were polymorphisms in the intron 3 of GH gene.
  Wenjin Zhu , Yongmei Su , Yanfang Liu , Xuemin Guan , Jing Ni and Jianhua Wu
  Growth of animal is largely regulated by Growth Hormone (GH). In this study the objective was to investigate variations in the fifth exon of GH gene and their polymorphism in 5 Chinese indigenous donkeys by PCR-SSCP and DNA Sequencing Methods. The results showed that there was a transversion at nucleotide position 1802 of GH gene in the 5th exon named as GH-exon5-G1802C which led to a conservative Lysine to Asparagine substitution at amino acid position 205. In detail, two different alleles, A and B were identified and three genotypes were observed, AA, AB and BB with the frequency distribution of allele B from 0.5962-0.7344 in analyzed populations. The genetic diversity analysis revealed that all PIC values were between 0.3140 and 0.3656, implying that this locus within GH gene possessed moderate genetic diversity in 5 Chinese indigenous donkeys. The χ2-test showed that GZ donkey was significant deviation, DZ donkey was no significant deviation from Hardy-Weinberg equilibrium, GL, JN and XJ donkey was not in agreement Hardy-Weinberg equilibrium. The results confirmed that there were polymorphisms in the exon 5 of GH gene.
  Wenjin Zhu , Yongmei Su , Yanfang Liu , Jing Ni and Jianhua Wu
  To reveal genetic diversity for the breed resource evaluation and utilization of Chinese indigenous donkey, polymorphisms in the third exon of GH gene was analyzed by PCR-SSCP and DNA sequencing methods in 16 donkey breeds (DZ, GZ, GL, JN, BH, LX, JM, QY, BY, HY, LZ, HB, GY, YY, TH and XJ). The results showed that there was a transition at nucleotide position 957 genomic DNA of GH gene, named as GH-exon3-G957A which led to a glutamic acid to lysine substitution at amino acid position 81. Two different alleles, A and B were identified and three genotypes were observed, AA, AB and BB with the frequency distribution of allele A from 0.6667-0.8333 in the analyzed populations. The genetic diversity analysis revealed that PIC values were between 0.346 and 0.491 except JM donkey (0.239), implying that this locus within GH gene possessed a moderate genetic diversity in Chinese indigenous donkeys. The χ2-test showed that LX, GY and TH donkeys were no significant deviation, GZ, JN, QY, HB and XJ donkey were significant deviation, DZ, GL, BH, JM, BY, HY, LZ and YY donkey were very significant deviation. The results confirmed that there were polymorphisms in the third exon of GH gene for the first time.
  Wenjin Zhu , Yongmei Su , Yanfang Liu , Jing Ni and Jianhua Wu
  To analysis the polymorphism of Yang Yuan donkey in China, the polymorphism of 12 microsatellites in 64 Yang Yuan donkeys were studied by PCR, polyacrylamide gel electrophoresis and silver staining. Number of alleles (Na), effective number of alleles (Ne), Polymorphism Information Content (PIC), observed Heterozygosity (Ho) and expected Heterozygosity (He) of each microsatellite were counted. And the Hardy-Weinberg equilibrium was tested as well.
 
 
 
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