Guan Weijun

Study on single nucleotide polymorphisms of H-FABP gene in sheep.

PCR-SSCP and DNA sequencing approaches were applied to assess the single nucleotide polymorphisms (SNPs) and analyze the genetic polymorphisms at partial exon 2 and intron 2 of H-FABP(Heart fatty acid-binding protein)gene, and five sheep populations that comprised of Small-Tailed Han sheep (SH, 48), Ningxia Tan sheep (Tan, 121), Tan x SH F1 (23), Poll Dorset (48) and Suffolk (24) sheep were screened in this study. The result showed: (1)four SNPs at 981(G/A), 1014(A/C) 1019(T/C) and 1058 (-/G ) and nine genotypes (AA, BB, CC, AB, AC, BC, AD, CD and BD) were detected using primer 2, the AA was the predominant genotype. A chi-square analysis suggested that the allele frequencies and genotype frequencies of H-FABP were in Hardy-Weinberg equilibrium except the Tan and Suffolk populations. Statistical analysis revealed a low polymorphism information content (PIC) in the Suffolk and Tan x SH F1 populations (PIC amp; 0.25) but an intermediate PIC in the remaining three populations (0.25 amp; PIC amp;0.50). It meant that the fragment of H-FABP had polymorphisms, which could be used as a candidate gene associated gene with phenotypic traits like intramuscular fat content in different sheep populations. (2)Three genotypes (HH, Hh and hh) determined by a SNP at 2407(T-C) were detected using primer 4. The genotype frequencies were in the order of HHHhhh. A chi-square analysis suggested that the allele frequencies and genotype frequencies of H-FABP were in Hardy-Weinberg equilibrium in the Tan and Poll Dorset populations, and the PIC values were low (PIC amp; 0.25). However, there was no polymorphisms in SH, Tan x SH F1 and Suffolk populations.

Establishment and characterization of a Chinese Game chicken embryo fibroblast bank

For preserving the genetic resources of the Chinese Game chicken and exploring a new method to preserve endangered animals, a Chinese Game chicken embryo fibroblast cell bank was established by the attachment culture method and the cryopreservation biotechniques. This bank includes 43 embryo samples and has stocks of 178 cryovials, with each containing 4.0×106 cells respectively. Quality assays and biological characteristics of the bank established showed that the cells cultured in vitro were all typical fibroblast and the cell population doubling time (PDT) was about 2 days. The cell viability before freezing was 97.37% ± 2.25% and 91.43% ± 3.22% after thawing. Tests for microbial contamination by bacteria, fungus, viruses and mycoplasmas were all negative. The frequency of cells having the diploid number of chromosomes (78) was 94.65% ± 3.29%. Isoenzyme analysis confirmed that there was no cross-contamitation in the culture. The fluorescence could be observed well-distributed in cytoplasm and nucleus except for some cryptomere vesicles at 24 h after transfection, and the expression efficiency of pEGFP-C1, pEGFP-N3, pEYFP-N1, pECFP-N1, pECFP-mito and pDsRed1-N1 were all between 9.5%∼34.6% at 24 h, 48 h and 72 h after transfection. This research provided both technical and theoretical support for preserving the genetic resources of other animals and poultry at the cell level.

Characterization of expression of α1-acid glycoprotein gene in Beijing fatty chicken (Gallus gallus).

: The specific expression of alpha1-AGP gene in eight different tissues of Beijing fatty chicken was investigated by RT-PCR. The full-length cDNA of alpha1-AGP was inserted into pEGFP-C1 multi-cloning sites to construct recombinant eukaryotic expression vector pEGFP-alpha1-AGP. The lipofectin method was used to transfect the pEGFP-alpha1-AGP into Beijing fatty chicken fibroblast cells. The open reading frame of Beijing fatty chicken alpha1-AGP gene was 612 base pairs in length, which was expressed higher in liver and lung than in muscle. This gene did not express in heart and kidney. The expression efficiency ranged from 31.3% to 47.6% in 24, 48, and 72 h after transformation. The green fluorescence mainly concentrated in the nucleus. With the increase of the expression of green fluorescence, granula was observed in the nucleus. RT-PCR and Western blotting analyses showed that pEGFP-alpha1-AGP had been integrated into the genome of Beijing chicken fibroblast cell with normal expression level. In optimized condition, there was no significant effect (P>0.05) on apoptosis ratio, positive cell shape, growth and reduplication state comparing with the control group. This research established the foundation for further function research of alpha1-AGP gene and application in transgenic animal cloning.

Testing the mutagenicity of five pesticides with micronucleus technique ofTradescantia paludosa

Applying the micronucleus techniqueTradescantia paludosa, the mutagenicity of five pesticides, Decis, Sumicid, Cypermethrin, Trichlorphon and Diazinon were tested. The testing results showed that 0.64%, 0.32%, 0.16%, 0.08% of Decis, 0.64%, 0.32%, 0.16%, 0.04% of Trichlorphon and 0.32%, 0.16%, 0.08%, 0.04% of Diazinon all had mutagenicity onTradescantia paludosa, but Sumicid and Cypermethrin had no such characteristic.

Research on the heredity and variation of the peruxidase isoenzyme of needles ofpinus koraiensis

Through the polyacrylamide gel electrophoresis analysis of thirty-two needle samples of ninty-years oldpinus koraiensis in Harbin, seven typies of zone and distribution of the zone frequency, and some objective fact of heredity and variation were determined. These informations can be used as reference materials in the research of heredity and variation ofpinus koraiensis.

Effect of low temperature on pollen vitality ofLarix olgensis

After the treatment of pollen oflarix olgensis at the different low temperature, the pollen vitality was observed by using fluorescent staining method. Several conclusions were obtained: 1) 0°C–3°C is the best temperature range for storing pollen ofLarix olgensis. 2) At the same temperature, humidity is the key factor influencing the pollen vitality. 3) Different clones shows different resistance to low temperature. Among the ten clones 2# clone shows the highest resistance.