Association between IL-10 genetic variations and cervical cancer susceptibility in a Chinese population.

We conducted an investigation into the role of the IL-10 polymorphisms -592A/C (rs1800872), -819C/T (rs1800871), and -1082A/G (rs1800896) in cervical cancer risk in a Chinese population. A case-control study was carried out, including 165 newly diagnosed cervical cancer patients and 165 control subjects. The polymerase chain reaction-restriction fragment length polymorphism method was used to genotype the three IL-10 variant loci. Using conditional logistic regression analysis, we observed that homozygous IL-10 -819C/T TT carriers were at significantly increased risk of cervical cancer compared to homozygous CC individuals, with an adjusted odds ratio (OR) of 2.23 and a 95% confidence interval (CI) of 1.16-4.30. Moreover, the CT+TT genotype was significantly associated with cervical cancer in comparison to the wild-type variant (OR = 1.69, 95%CI = 1.04-2.76; P = 0.03). In conclusion, our study suggests that the IL-10 -819C/T genetic variation may contribute to cervical cancer risk in the Chinese population examined.

Isolation and characterization of polymorphic microsatellite markers for blue fox (Alopex lagopus).

The blue fox, belonging to the family Canidae, is a coat color variant of the native arctic fox (Alopex lagopus). To date, microsatellite loci in blue fox are typically amplified using canine simple sequence repeat primers. In the present study, we constructed an (AC)n enrichment library, and isolated and identified 17 polymorphic microsatellite markers for blue fox. The number of alleles per locus is from two to seven based on 24 examined individuals. The expected and observed heterozygosities were in the range of 0.3112 to 0.8236 and 0.2917 to 0.8750, respectively. The polymorphic information content per locus ranged from 0.2583 to 0.8022. These polymorphic markers can be useful for future population genetic studies of both farmed blue foxes and wild arctic foxes.

De novo assembly and characterization of farmed blue fox (Alopex lagopus) global transcriptome using Illumina paired-end sequencing.

The blue fox (Alopex lagopus), a coat-color variant of the Arctic fox, is a domesticated fur-bearing mammal. In the present study, transcriptome data generated from a pool of nine different tissues were obtained with Illumina HiSeq2500 paired-end sequencing technology. After filtering from raw reads, 32,358,290 clean reads were assembled into 161,269 transcripts and 97,252 unigenes by the Trinity fragment assembly software. Of the assembled unigenes, 37,967 were annotated in the National Center for Biotechnology Information (NCBI) Non-Redundant (NR) protein database and 26,264 in the Swiss-Prot database. Among the annotated unigenes, 24,839 and 24,267 were assigned using the Gene Ontology (GO) and euKaryotic Orthologous Groups (KOG) databases, respectively. Altogether, 17,057 unigenes were mapped onto 227 pathways using the Kyoto Encyclopedia of Genes and Genomes database. In addition, 6394 simple sequence repeats were identified by examining 12,965 unigenes (>1 kb), which could contribute to the development of molecular markers. This study generated transcriptome data for the blue fox that will promote further progress in expression profiling studies, and provide a good annotation basis for genomic studies.

Identification of cDNA sequences and alternative splicing patterns of canine AMEL genes (AMELX and AMELY).

Amelogenin is a major protein of the developing enamel matrix. There are two amelogenin genes (AMELX and AMELY) located on the X and Y chromosomes, respectively, in dogs. In the present study, we characterized full-length cDNAs and alternative splicing patterns of the AMEL genes in the tooth tissue of a dog by 5'- and 3'-rapid amplification of cDNA ends and AMEL-specific RT-PCR. Sequence analysis revealed that the coding regions of AMELX and AMELY were 579 and 576 bp (accession Nos. KP244310 and KP244311), respectively. The coding sequence of AMELX had 95.1% identity to that of AMELY. The AMEL genes on X and Y chromosomes were both expressed in developing tooth tissue. Eight different alternatively spliced transcripts were identified, five from AMELX and three from AMELY.

Cloning and identification of the ASIP gene in Chinese raccoon dog (Nyctereutes procyonoides procyonoides).

The quantity, quality, and distribution of eumelanin and pheomelanin determine a wide variety of coat colors in animals. Three coat color variants exist in farmed wild-type Chinese raccoon dog (Nyctereutes procyonoides procyonoides), which is an important fur-bearing animal species. The ASIP gene is an important candidate gene for coat color variation in some species. In this study, the complete cDNA sequences of ASIP were amplified from a wild-type Chinese raccoon dog. Sequence analysis revealed the coding region of ASIP in Chinese raccoon dog to be 396-bp in length and two transcripts (accession Nos. KT224450 and KT224451) were identified due to the alternative use of exon 1 (1A and 1C). However, the alternative splicing pattern and the coding sequence of ASIP in three types of coat color variants were the same as those identified in the wild-type individual. Based on the results obtained in this study, we can exclude a role for alternative splicing of exon 1 and the coding sequence of ASIP in coat color variation in Chinese raccoon dog.

Isolation and characterization of novel microsatellite markers from the sika deer (Cervus nippon) genome.

Microsatellite markers are widely and evenly distributed, and are highly polymorphic. Rapid and convenient detection through automated analysis means that microsatellite markers are widely used in the construction of plant and animal genetic maps, in quantitative trait loci localization, marker-assisted selection, identification of genetic relationships, and genetic diversity and phylogenetic tree construction. However, few microsatellite markers remain to be isolated. We used streptavidin magnetic beads to affinity-capture and construct a (CA)n microsatellite DNA-enriched library from sika deer. We selected sequences containing more than six repeats to design primers. Clear bands were selected, which were amplified using non-specific primers following PCR amplification to screen polymorphisms in a group of 65 unrelated sika deer. The positive clone rate reached 82.9% by constructing the enriched library, and we then selected positive clones for sequencing. There were 395 sequences with CA repeats, and the CA repeat number was 4-105. We selected sequences containing more than six repeats to design primers, of which 297 pairs were designed. We next selected clear bands and used non-specific primers to amplify following PCR amplification. In total, 245 pairs of primers were screened. We then selected 50 pairs of primers to randomly screen for polymorphisms. We detected 47 polymorphic and 3 monomorphic loci in 65 unrelated sika deer. These newly isolated and characterized microsatellite loci can be used to construct genetic maps and for lineage testing in deer. In addition, they can be used for comparative genomics between Cervidae species.

Development of novel polymorphic microsatellite markers for the silver fox (Vulpes vulpes).

The silver fox (Vulpes vulpes), a coat color variant of the red fox, is one of the most important fur-bearing animals. To date, development of microsatellite loci for the silver fox has been limited and mainly based on cross-amplification by using canine SSR primers. In this study, 28 polymorphic microsatellite markers were isolated and identified for silver fox through the construction and screening of an (AC)n-enriched library. The number of alleles per locus ranged from 2 to 8 based on 48 individuals tested. The expected and observed hetero- zygosity and polymorphism information content per locus ranged from 0.2544 to 0.859, 0.2083 to 0.7917, and 0.2181 to 0.821, respectively. The polymorphic markers presented in this study may be useful for future analysis of the genetic diversity and population structure of farmed silver fox and wild red fox.

Cloning and association analysis of KIT and EDNRB polymorphisms with dominant white coat color in the Chinese raccoon dog (Nyctereutes procyonoides procyonoides).

The Chinese raccoon dog (Nyctereutes procyonoides procyonoides) is one of the most important fur-bearing animal species. The dominant white individual, a coat color variant in farmed Chinese raccoon dog, shows a completely white phenotype over the entire body. The KIT and EDNRB genes have been reported to be associated with the dominant white coat color in some mammalian species. In the present study, the full-length coding sequences of KIT and EDNRB were amplified from a dominant white and a wild-type Chinese raccoon dog. Sequence analysis revealed that the coding region of KIT and EDNRB in Chinese raccoon dog was 2919 and 1332 base pairs in length (accession No. KM083121 and KM083122), respectively, and 2 single-nucleotide polymorphisms (SNPs; c.600C>T and c.967G>A) in KIT and 1 SNP (c.259A>C) in EDNRB was found only in the dominant white individual. An alternative splicing site at the boundary of 4 and 5 of the KIT gene was identified in both individuals. We further investigated the association between the 3 SNPs of KIT and EDNRB and dominant white coat color by genotyping 18 individuals. We found no association between these SNPs and dominant white coat color. Based on these results, we can exclude the coding regions of the KIT and EDNRB genes as determinants of the dominant white coat color in Chinese raccoon dog.

Development and characterization of polymorphic microsatellite markers for Chinese raccoon dog (Nyctereutes procyonoides procyonoides).

Chinese raccoon dog (Nyctereutes procyonoides procyonoides) is one of the most important fur-bearing animal species. Information about the genetic background of farmed Chinese raccoon dogs is limited. In this study, 17 polymorphic microsatellite markers were isolated and identified from an (AC)n-microsatellite-enriched library of Chinese raccoon dogs. The number of alleles per locus ranged from 2 to 8 based on 48 individuals tested. The expected and observed heterozygosity and polymorphism information content per locus ranged from 0.383 to 0.8378, 0.3200 to 0.8696, and 0.3047 to 0.7947, respectively. Cross-species amplification of these loci in 2 other Canidae species indicated that 9 and 11 of these loci could also be amplified successfully in the arctic and silver fox, respectively. These microsatellite loci developed in the present report will provide useful tools for population genetic studies, individual identification, and phylogenetic analysis in the Chinese raccoon dog and other Canidae species.