Wu ChangXin

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

Myostatin is a negative regulator of skeletal muscle mass. The present study cloned the 5′ regulatory region of porcine myostatin gene, screened its polymorphisms and analyzed their associations with early growth traits in Yorkshire pigs. The results indicated that a fragment length polymorphism and a polymorphism concerning two nucleotide changes exist in the 5′ regulatory region of porcine myostatin gene. At sites 435 and 447, allele A and allele B have the haplotypes of A-G and G-A, respectively. The allelic frequency of B is 0.475 in Yorkshire pigs. No homozygous BB genotype was detected in 9 Laiwu Black pigs. Allele B was found to have positive effect on body weight 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), respectively. The additive effect of allele B on BW21, BW28, BW70, ADG1, ADG2 and ADG3 was 0.596±0.205 kg (P=0.0041), 0.498±0.200 kg (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) and 37.00±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 AA and AB.

Sequencing and alignment of mitochondrial genomes of Tibetan chicken and two lowland chicken breeds

Tibetan chicken lives in high-altitude area and has adapted well to hypoxia genetically. Shouguang chicken and Silky chicken are both lowland chicken breeds. In the present study, the complete mitochondrial genome sequences of the three chicken breeds were all sequenced. The results showed that the mitochondrial DNAs (mtDNAs) of Shouguang chicken and Silky chicken consist of 16784 bp and 16785 bp respectively, and Tibetan chicken mitochondrial genome varies from 16784 bp to 16786 bp. After sequence analysis, 120 mutations, including 4 single nucleotide polymorphisms (SNPs) in tRNA genes, 9 SNPs and 1 insertion in rRNA genes, 38 SNPs and 1 deletion in D-LOOP, 66 SNPs in protein-coding genes, were found. This work will provide clues for the future study on the association between mitochondrial genes and the adaptation to hypoxia.

Effects of different states of sheep fetal fibroblasts as donor cells on the early development in vitro of reconstructed sheep embryos

To investigate the effects of different states of donor cells on the development of reconstructed sheep embryos, we designed five treatments of donor cells, including cell passage, cell size, serum starvation, colchicine treatment and gene transfection. Results are as follows: (I) Compared with 16–18 passage cells, the morula/blastocyst rate of 5–7 passage cells as donor nuclei was significantly higher (17.3% vs. 4.9%, P<0.05), suggesting the advantage of short-time cultured cells in supporting the development of reconstructed embryos. (II) The mourla/blastocyst rate of reconstructed embryos derived from medium cells (15–25 μm) as donor nuclei was higher than that from large cells (25–33 μm) and small cells (8–15 μm)(20.0% vs. 8.0%, 9.7%), indicating that reconstructed embryos from medium cells had a greater potentiality to develop into morula/blastocysts than those from small or large ones. (III) The morula/blastocyst rate of reconstructed embryos from donor cells of SS (serum starvation) was lower than that from donor cells of NSS (non-serum starvation), but no significant difference was detected between SS and NSS(11.8% vs. 18.6%, P>0.05). (IV) Fetal fibroblasts treated with 0.05 μmol/L colchicine exhibited a higher morula/blastocyst rate of reconstructed embryos than those treated with 0.10 μmol/L colchicine and untreated ones (27.5% vs. 12.1%, 17.1%), however, no significant difference among the three treatments was detected (P>0.05). (V) The morula/blastocyst rate of reconstructed embryos from fetal fibroblasts transfected with GFP gene only was 3.1%, significantly lower than that from non-transgenic cells (3.1% vs. 20.4%, P<0.05). In conclusion, our results demonstrated that fetal fibroblasts of fewer passages, medium size could ensure a higher morula/blastocyst rate of reconstructed embryos. Serum starvation of donor cells might be unnecessary to the development of reconstructed embryos. Donor cells treated with 0.05 μmol/L colchicine could facilitate the development of reconstructed embryos. Additionally, as cells transfected with GFP gene were used as donor nuclei, adverse effect on the development of reconstructed embryos was observed. Therefore, the developmental efficiency of reconstructed embryos could be improved if proper treatments to donor cells were used.

Single nucleotide polymorphisms in chicken lmbr1 gene were associated with chicken growth and carcass traits

Lmbr1 is the key candidate gene controlling vertebrate limb development, but its effects on animal growth and carcass traits have never been reported. In this experiment, lmbr1 was taken as the candidate gene affecting chicken growth and carcass traits. T/C and G/A mutations located in exon 16 and one A/C mutation located in intron 5 of chicken lmbr1 were detected from Silky, White Plymouth Rock broilers and their F2 crossing chickens by PCR-SSCP and sequencing methods. The analysis of variance (ANOVA) results suggests that T/C polymorphism of exon 16 had significant association with eviscerated yield rate (EYR), gizzard rate (GR), shank and claw rate (SCR) and shank girth (SG); A/C polymorphism of intron 5 was significantly associated with SCR, liver rate and head-neck weight (HNW), while both sites had no significant association with other growth and carcass traits. These results demonstrate that lmbr1 gene could be a genetic locus or linked to a major gene significantly affecting these growth and carcass traits in chicken.

An association between tyrosinase gene single nucleotide polymorphisms and melanin distribution in chicken

Tyrosinase (TYR) is a key enzyme of melanin biosynthesis. Single-strand conformation polymorphism (SSCP) analysis was applied to detect the single nucleotide polymorphisms (SNPs) in the upstream regulating region from -641 to -2125 bp of the TYR gene. Three SNPs were found in this region. Correlations were obtained between genotypes of the SNP sites and pigment traits in chicken parental and F 2 generations of the Chinese Agricultural University (CAU) resource population. The chi-square test indicated that these mutations were significantly related to shank and body skin colours.