ABSTRACT
Reproduction is a complex physiological process that is regulated by multiple genes and pathways. Compared with studies of common livestock, fewer studies of genes related to the fertility of rabbits (Oryctolagus cuniculus) have been reported, and the molecular mechanism of their high productivity is still poorly understood. To identify candidate genes associated with development and prolificacy in rabbits, we analyzed gene expression differences among the ovaries of mature Californian rabbit (LC), and mature (HH) and immature Harbin white rabbit (IH) using digital gene expression technology. We detected 885 and 321 genes that were significantly differentially expressed in comparisons between HH/IH and HH/LC, respectively. The functions of the differentially expressed genes (DEGs) were determined by GO classification and KEGG pathway analysis. The results suggest that most of the DEGs between the mature and immature developmental stages were predominantly associated with DNA replication, cell cycle, and progesterone-mediated oocyte maturation, and most were up-regulated in the IH group compared with the HH group. The DEGs involved in disparate fecundities between HH and LC were associated with reproduction, fructose and mannose metabolism, steroid hormone biosynthesis, and pyruvate metabolism. Our results will contribute to a better understanding of changes in the regulatory network in ovary at different developmental stages and in different fertility of rabbit.
Subject(s)
Gene Expression Profiling/methods , Oogenesis/genetics , Reproduction/genetics , Transcriptome/genetics , Animals , Female , Fertility/genetics , Gene Expression Regulation, Developmental/genetics , Gene Regulatory Networks/genetics , High-Throughput Nucleotide Sequencing , Microarray Analysis , Ovary/metabolism , RabbitsABSTRACT
Zygote arrest 1 (Zar1) is an oocyte-specific maternal-effect gene. Previous studies indicate that Zar1 plays important role in early embryo development, but little is known about its function in rabbit. The objectives of this study were to clone the New Zealand white rabbit Zar1 gene and to investigate its expression in various organs in groups of animals with different reproductive traits.We obtained a 709-bp Zar1 cDNA fragment consisting of an 8-bp exon 1, 161-bp exon 2, 75-bp exon 3, 271-bp exon 4 and 194-bp 3'sequences. The rabbit Zar1 nucleotide sequence showed per cent identities of 91, 88, 88, 87, 86, 87, 76 and 82% with Zar1 orthologues in human, cattle, sheep, pig, mouse, rat, zebrafish and Xenopus laevis, respectively, indicating a high homology with other species and evolutionary conservation. Quantitative real-time polymerase chain reaction analyses revealed nonoocyte-specific Zar1 expression, with expression in spleen, lung, ovary, uterus, heart, liver and kidney. The expression level was highest in the lung. This study may lay the theoretical foundation for the study of ZAR1's biological function.
Subject(s)
Cloning, Molecular , Egg Proteins/genetics , Gene Expression , Animals , DNA Methylation , DNA, Complementary , Egg Proteins/chemistry , Egg Proteins/metabolism , Organ Specificity/genetics , RNA, Messenger/genetics , Rabbits , Real-Time Polymerase Chain ReactionABSTRACT
A novel high-throughput method was established for rapid screening of a large numbers of Aspergillus fumigatus (A. fumigatus) mutants with high chitosanase production under acidic culture condition by exploiting the fact that iodine can be used as the indicator to stain chitosan but is ineffective for chitooligosaccharides. The mutant population was generated by irradiating A. fumigatus CICC 2434 with Co(60)-γ rays. Mutants were cultured on acidic plates containing colloidal chitosan and preliminary screened according to diameter of haloes formed around colonies. Then, chitosanase production of the isolates were verified by dinitrosalicylic acid assay. Lastly, molecular masses on enzymolysis products of isolated mutants were rapidly compared by aniline blue plate assay. Using this method, the mutant strain Co-8 was selected, which had chitosanase activity of 24.87 U/mL (increased by 369.2 % as compared to that of its parental strain).Taking together, the method is easy, efficient and particularly suited to rapid screen acidophilic fungal strains with high chitosanase-production.
Subject(s)
Acids/chemistry , Aspergillus fumigatus/enzymology , Aspergillus fumigatus/isolation & purification , Glycoside Hydrolases/metabolism , High-Throughput Screening Assays/methods , Aspergillus fumigatus/genetics , Aspergillus fumigatus/radiation effects , Culture Media/chemistry , Glycoside Hydrolases/genetics , Iodine/chemistry , Molecular Weight , MutationABSTRACT
Bone morphological protein 7 (BMP7) has been proposed to be an osteoinductive protein. Recent data have shown that BMP7 also plays a crucial role in the growth and development, and physiological function of reproductive system. To date, studies have shown an association between the BMP gene family and reproduction in many populations, but few studies have completely described this association in sow. In the present study, three sow breeds were screened out to investigate the genetic effects of the BMP7 gene. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing, three single nucleotide polymorphisms (SNPs) (g.35161T>C, g.35175T>C and g.35216C>T) were identified in intron 2 of the BMP7 gene. Associations between the three SNPs and the number of piglet born alive (NBA), litter weight at birth (LBW), total number of piglet born (TNB) and litter weight at 21 days were analyzed using association analysis. Among the three SNPs, g.35161T>C was significantly associated with NBA and LBW (p<0.05), and the litter weight at 21 days (p<0.01). These results suggest that g.35161T>C is a potential candidate gene locus for litter size traits and the BMP7 gene might be associated with the quantitative trait locus (QTL) controlling the litter size. These data will provide a background for more extensive characterization of the BMP7 gene.
