Biomedical subjects

Chang-Xin Wu

Publications and source records attributed to Chang-Xin Wu.

At least 19 recordsLinked to original sources

PubMed2006

Effects of oocyte activation and sperm preparation on the development of porcine embryos derived from in vitro-matured oocytes and intracytoplasmic sperm injection.

The objective was to determine the effects of various methods of oocyte activation and sperm pretreatment on development of porcine embryos derived from in vitro-matured oocytes and intracytoplasmic sperm injection (ICSI). The second polar body was extruded in the majority (>78.4%) of in vitro-matured (IVM) oocytes 4h after electrical pulse activation. In embryos generated by ICSI and sham-ICSI, a combination of an electrical pulse, with various chemical activators 4 h later, improved (P < 0.05) blastocyst formation rate compared to activation only with a pulse. Treatment with 6-dimethylaminopurine (DMAP) after electrical activation significantly increased the oocyte activation rate. The effects of exposure of sperm to repeated freeze-thaw cycles (without cryoprotectant) on oocyte activation and the effects of sperm pre-incubated with dithiothreitol (DTT) or Triton X-100 on early embryo development were also examined. Blastocyst formation rates after ICSI did not differ between motile sperm and those rendered immotile by one-time freezing and thawing without cryoprotectant. However, sperm rendered immotile by three cycles of freezing/thawing without cryoprotectant had a significantly lower blastocyst formation rate. Although oocytes injected with sperm pre-incubated with Triton X-100 had a higher normal fertilization rate than those pre-incubated with DTT or one-time frozen/thawed sperm, rates of blastocyst formation and cell numbers were similar among the three groups. In conclusion, various methods of oocyte activation and sperm preparation significantly affected the developmental capacity of early porcine embryos derived from IVM and ICSI.

Animals↗

PubMed2006

Analysis of expressed sequence tags from skeletal muscle-specific cDNA library of Chinese native Xiang pig.

A Longissimus Dorsi muscle cDNA library of Xiang Pig was constructed, and 131 randomly isolated clones were sequenced in this study. The results of bioinformatics analysis showed that 131 ESTs represented 109 unique clones sequences, of which 99 showed homology to previously identified genes in humans or other mammals, 3 matched other uncharacterized expressed sequence tags (ESTs), and 7 showed no significant matches to sequences already present in DNA databases. No protein matches were found for 10 ESTs. Functional analysis of the ESTs showed that a considerable proportion of them encoded proteins involved in gene/protein expression (45.46%). Other classes included genes involved in metabolism (10.10%), cell structure/motility (10.10%), cell/organism defense (5.05%), cell signaling/communication (2.02%), and cell division (0.0%). Unclassified genes constituted the remaining 27.27%. This study reported the results of the first gene expression profile analysis of Chinese native Xiang Pig skeletal muscle cells, thereby greatly facilitating the functional study of candidate genes involved in muscle growth as well as in the improvement of meat quality in domestic pigs.

Animals↗

PubMed2006

[Identification of a common deletion in different curly mutants in Drosophila melanogaster].

Curly is a easily distinguishable dominant mutant wing character. The Cy mutation is the most commonly used dominant marker for the second chromosome balancers in Drosophila melanogaster, but little is known about the Cy gene. Based on known genomic and cytogenetic information, a 102 bp deletion which is located between the Genes synaptotagmin (syt) and Activin Like Protein at 23B(Alp23B) on the Drosophila melanogaster genome (release 4) had been found to be commonly contained on Cy chromosome in three different curly strains. Meanwhile, when using the deletion as a DNA marker, the result suggested that Cy homozygote be lethal in embryo period. These results will provide some helpful information to investigate molecular mechanism of curly wings in the further study.

Animals↗

PubMed2006

[Genetic analysis of feather color and shank color traits based on F-2 resource population in Tibetan chicken].

Tibetan chickens were crossed reciprocally with White Leghorn and Shou-Guang chicken respectively, and inter se matings in F1 were carried out to generate the F2 population. Feather color and shank color appearance and segregation ratio in F1 and F2 were observed. Results indicate that white feather of White Leghorn chicken and black feather of Shou-Guang chicken exhibit complete dominant heredity to hemp feather of Tibetan chicken. Hemp feather is determined by two or more alleles. Only when these two or more alleles are concurrently present, will hemp feather then be displayed. The Id/id allele that determines shank color demonstrates sex-linked inheritance, and the recessive id is expressed gradually in the homozygotes. We confirm that genotype of shank color in White Leghorn rooster used in this study is the dominant homozygote.

Animal Structures↗

PubMed2005

[Sequencing analysis of the control region and intron1 region and characterization of two novel microsatellites in obese gene of pig].

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.

5' Untranslated Regions↗

PubMed2004

[Identification and analysis of a novel microsatellite marker flanking porcine myostatin gene (MSTN)].

In animal breeding, microsatellite marker plays an important role in constructing genetic maps, QTL mapping and function analysis of structural genes. Myostatin, also known as GDF8, is a negative regulator of skeletal muscle mass and, in swine, it is evidenced to be related to birth weight and average daily gain from 60 kg to 100 kg of body weight. In present study, by subcloning and sequencing,we identified a novel microsatellite marker which is useful for fine QTL mapping for meat traits. A BAC clone containing porcine MSTN was extracted and digested with EcoR I to recover the fragment of > 4 kb for subcloning in pGEM-3zf (+). Sequencing and alignment results showed that this subcloned fragment was not from porcine MSTN, but included a tandem repeat of (TG) 13, which is a novel microsatellite marker (GenBank accession number: AF454400) flanking MSTN. To exclude its vector origin we designed specific primers flanking this marker and successfully amplified this fragment from porcine genome. Through a pedigree analysis of a double-muscled Yorshire strain, we found that it is inherited in a co-dominant manner. We also checked the gene frequencies of this locus in 381 unrelated individuals of 7 pig breeds, namely Laiwu,Landrace, Yorkshire,Duroc, Peterian, Min and Erhualian. Only two alleles were detected, the repeating number of which are 13 (allele A) and 19 (allele B) respectively, which indicated that it is a low poly morphic microsatellite marker. In addition, the frequencies of the two alleles are different between the two types of pig breeds, while allele A is dominant in Chinese local breeds, allele B is dominant in imported breeds. Alignment with AY208121 indicate that this locus is located 42 kb downstream of porcine MSTN. We speculate that this microsatellite DNA is an important marker both in fine QTL mapping for meat traits and in the expression study of porcine MSTN.

Animals↗

PubMed2004

[Construction and identification on enriched microsatellite library from yak genome].

We constructed the first microsatellite-enriched library of yak according to the strong affinity between biotin and streptavidin. The method included ligation of 300 approximately 1 000 bp enzyme-digested fragments and adaptors, affinity capture of microsatellite repeat using biotinylated oligoprobe ((CA)12, (CCG) 8, (CAG)8, (TTTC) 8) attached to streptavidin-coated magnetic beads, PCR amplification using the 21-mer adaptor oligonucleotide as primer to obtain double-stranded targeted fragments, religated into pMD18-T vector and transformed to DH5alpha. The results of sequencing showed that 37 of 48 readable sequences contained microsatellites indicating a high degree of microsatellite enrichment. The new polymorphic microsatellite markers we have identified and characterized will contribute to the yak genetic linkage mapping, molecular evolution and phylogenetic studies, marker assistant selection and QTLs location of yak main economic traits.

Animals↗

PubMed2004

[A new method for phylogenic analysis].

With ESTs from porcine fatty tissue and cDNA sequences from human, bovine and mouse in non-reduncdant database and dbEST in GenBank,we sampled cDNA sequences of 70 function-known genes in four species on the base of randomly sampling method, analyzed the mutation pattern of 70 x 150 bp linking sequences between them, and established an integrated phylogenic analysis method. The results showed that 391 single bases mutations were found in 70 x 150 bp linking sequences alignment among four species. The number of mutation bases between them were greatly exceeded the 1/1000 predicted in the human genome analysis. C/T(T/C) and A/G (G/A) transitions were the major types of single base mutation. The genetic relationship between pig and bovine who are both Artiodactylous is the nearest, the next is human, and the farthest is mouse. The differentiation sequence taken place in four species from the same ancestor is that mouse is the earliest one, and the latter human, and pig and bovine are the latest.

Animals↗

PubMed2004

[Chromosomal mapping of chicken MC4R using a radiation hybrid panel and the comparative analysis of the gene homologous regions between chicken and human chromosome].

Mutations of the melanocortin-4 receptor (MC4R) are associated with hyperphagia, obesity, and accelerated longitudinal growth in pig, mice and human. However, little is known about the functions of this gene in chicken. To map the MC4R gene in Chicken chromosome, we used a 6000 rads chicken-hamster radiation hybrid panel (ChikenRH6). PCR of samples from the ChikenRH6 revealed the location of the MC4R gene to be nearby markers MCW0062, BCL2 and OVY on chromosome 2q12. Five markers were placed into a single linkage group based on two-point analysis with a LOD score of greater than 5. At the same time, the MC4R gene was selected as marker to compare DNA sequence between chicken and human chromosome. The result shows there are the same homologous regions between chicken chromosome 2 (GGA2) and human chromosome 18 (HSA18), and we found that the genes BCL2 and obesity are located in the near regions of MC4R on human chromosome 18. So we can reduce that the chicken MC4R gene maybe there are the same functions with the human MC4R gene. Overall, this work reveals widespread chromosome rearrangements of MC4R between chicken and human genomes, and mappings the chicken MC4R gene on 2q12 by a RH panel.

Animals↗

PubMed2004

[MC1R--the key gene in mammalian melanin synthesis].

The study of the molecular regulation mechanism of melanin synthesis during animal development has become a new focus recently . The synthesis and production of melanin during animal development are regulated by many genes. This paper summarized the molecular function mechanism of melanocortin-1-receptor (MC1R) gene and the relationship between the consequences of polymorphic variation of the gene and melanin traits, in addition to summarized the identification and mutation of MC1R gene in birds. Furthermore, the melanin synthesis mechanism in birds is also discussed here.

Adrenocorticotropic Hormone↗

PubMed2003

Primary factors affecting the evaluation accuracy of animal model MBLUP.

Marker-assisted best linear unbiased prediction (MBLUP) is one important method of implementing marker-assisted selection (MAS). The computer stochastic simulation method was used to study the effects of heritability of trait, QTL variance and map distance between two adjacent markers on the evaluation accuracy of animal model MBLUP. With the higher heritability, the smaller QTL variance and the shorter map distance between two adjacent markers, the evaluation accuracy of animal model MBLUP would be increased. On the contrary, the evaluation accuracy of animal model MBLUP would be decreased when the heritability was lower, QTL variance was larger and map distance between two adjacent markers was longer.

Animals↗

PubMed2003

Effects of marker-assisted selection under different initial frequency of QTL favorable allele.

A ten-generation continuous selection experiment on a single trait in a closed population was carried out by stochastic simulation. It assumed that the trait was controlled by polygenes and a single autosomal bi-allelic marked quantitative trait locus (QTL). Individual breeding values were estimated through animal model marker-assisted best linear unbiased prediction (MB-LUP), and breeding animals were selected according to their breeding values. The effects of three levels initial frequencies of QTL favorable allele on marker-assisted selection (MAS) were studied. The results showed that it would gain higher genetic response when implementing MAS on those traits with lower initial frequency of QTL favorable allele. When the initial frequency of QTL favorable allele was low, although the generation number required for fixing QTL favorable allele in the population would be lengthened, its frequency would be increased faster. However, the initial frequency of QTL favorable allele had little effects on the inbreeding rates.

Algorithms↗

PubMed2003

[Preliminary analysis on China Agricultural University chicken resource population based on genomic scanning].

Combining the technique of multiplex-PCR and the fluorescent semi-automated detection, a large-scale genome scanning was performed for 440 chickens, which was derived from China Agricultural University chicken resource families, within three generations. Fifty-five microsatellite markers were analyzed for this study. Those 55 microsatellite loci accorded with the characters of Mendelian co-inheritance. The heterozygosities ranged from zero to 0.89, with 72% of loci having a heterozygosity of more than 0.60. The polymorphism information content (PIC) ranged from 0 to 0.85, in which 70% of those loci had a PIC of more than 0.50 but their distribution varied in line A and line C. The allele frequency was significantly different between line A and line C at most loci (P < 0.01). At the same time, gene accordance inclination was found in line C. The Nei population resemble coefficient and standard genetic distance were 0.1002 and 0.8928.

Animals↗

PubMed2002

[Single nucleotide polymorphism analysis in chicken insulin-like growth factor-II gene and its associations with growth and carcass traits].

In this experiment, F2 chicken derived from Broilers crossing with Silky was used to study the effect of insulin-like growth factor-II gene on growth and carcass traits. The partial gene was amplified by two pairs of primers, and single nucleotide polymorphism (SNPs) was detected by the technique of restriction fragment length polymorphism (RFLP), and then confirmed by DNA sequencing. The mutation was found in the exon-2 of the gene, and can be clarified by cutting of restriction enzyme Aci-I. The result of least square analysis showed the gene was significantly related with growth and carcass traits. It implied that the insulin-like growth factor-II gene could be a genetic locus or linked to a major gene affecting greatly the growth and carcass traits in chicken.

Animals↗

PubMed2002

[Cloning and sequencing of fatty acid binding proteins gene in chicken].

A pair of primers was designed according to the sequence of mammal fatty acid binding protein (FABP) gene, then PCR amplified to chicken genome. After the product of PCR was cloned and sequenced, homologous comparison was done among porcine heart fatty acid binding protein gene and porcine adipocyte fatty acid binding protein gene. The result showed that the sequence of chicken FABP gene had 68% and 75% homology with porcine H-FABP and A-FABP gene respectively, and had 75% homology with porcine AFABP on amino acid level. The result of Northern showed that the gene only expressed in fat tissues.

Adipose Tissue↗

PubMed2002

[Relationship of T-->A mutation in the promoter region of myostatin gene with growth traits in swine].

A T-->A mutation in the promoter region of porcine myostatin (MSTN) gene has been identified in previous work. Associations of the myostatin genotypes with growth traits are unknown in swine. The present study attempts to analyze the relationship of the mutation with the growth traits which included body weight at 60 d (BW60), average daily gain from 25 kg to 60 kg(ADG1), average daily gain from 60 kg to 100 kg (ADG2) and average daily gain from 25 kg to 100 kg (ADG). Data from 165, 275, 276 and 276 unrelated individuals respectively were collected from three different swine breeding companies. Detections of the mutation were carried out by PCR-RFLP approach. The effect of MSTN genotypes (TT and TA) on growth traits was estimated by GLM procedure. The results showed that for ADG2, individuals with TA genotype were higher than those of TT genotype (P = 0.052), indicating a positive effect for A allele. For BW60, ADG1 and ADG, the effect of porcine MSTN genotype was non-significant (P > 0.1). Studies are still necessary for examining the effects in "double-muscled" pigs.

Alleles↗

PubMed2002

[Polymorphism analysis of the exon 2 of SLA-DQB gene in Xiao Meishan, Zhong Meishan and Yorkshire pigs with PCR-RFLP].

SLA is an important genetic region which influences disease resistance in pigs, and the gene family is one of important candidates for disease resistance and susceptibility. In present research the exon 2 of SLA-DQB gene was amplified and a uniform fragment of 273 bp was obtained in Xiao Meishan, Zhong Meishan and Yorkshire Pigs. The 273 bp PCR product was digested with restriction endonuclease Rsa I and genetic Polymorphism was investigated with PCR-RFLP (restriction fragment length polymorphism). Polymorphic sites were detected at base position 11, 94 and 124 of the exon 2 of SLA-DQB gene. There were two kinds of genotypes in Xiao Meishan pigs, 246 bp/27 bp(AA) and 132 bp/84 bp/30 bp/27 bp(BB), the frequencies of genotype AA and BB are 0.571, and 0.439 respectively. Four kinds of genotypes were found in Zhong Meishan pigs, 246 bp/27 bp (AA), 132 bp/84 bp/30 bp/27 bp(BB), 132 bp/114 bp/27 bp(CC) and 246 bp/132 bp/84 bp/30 bp/27 bp(AB), their frequencies are 0.037, 0.593, 0.037 and 0.333 respectively. Five kinds of genotypes were found in Yorkshire pigs, 246 bp/27 bp(AA), 246 bp/132 bp/114 bp/27 bp(AC), 132 bp/114 bp/27 bp(CC), 246 bp/132 bp/84 bp/30 bp/27 bp(AB) and 273 bp/246 bp/27 bp(AD), the frequencies of five genotypes are 0.467, 0.300, 0.100, 0.100 and 0.033 respectively. Statistical results indicated that there is no favorable allele in Xiao Meishan pig breed; B is the favorable allele in Zhong Meishan and its gene frequency is 0.759; while in Yorkshire pigs A is the favorable allele and its gene frequency is 0.683. Four alleles (A, B, C and D) were found in SLA-DQB locus among three pig breeds; The results of chi 2 showed that four alleles of this locus fit with Hardy-Weinberg equilibrium. The research showed that conservation of Chinese local pig breeds is necessary in order to protect our rich genetic resources and make use of them in the future.

Alleles↗

PubMed2002

[Analysis of succeed percentage on sequencing ESTs and construction of porcine muscle cDNA library].

Using Longissimus Dorsi muscle as material and Lambda ZAP II as Vector, Xiang Pig Longissimus Dorsi muscle cDNA library has been constructed in our study. The results showed that the titration of the library was 3.4 x 10(7) pfu/ml, the recombinant percentage was 94%, and the fragment length of inserted average cDNA were 1.5 kb. The study pointed out that the more than 30 T insertion is the major factor for low percentage if sequencing the 3'-end.

English Abstract↗