The long-tailed goral (Naemorhedus caudatus) is an endangered animal species in all its habitats worldwide, including South Korea. The imbalanced sex ratio in fragmented habitats is closely associated with extinction. Therefore, sex identification using wild animal samples would be necessary. However, only a few studies have been reported about the sex identification of gorals. In this study, we thus aimed at comparing the efficiency of sex identification using various goral sample types as templates and the amelogenin (AMEL) and DEAD-box polypeptide 3 (DDX3) genes as target sequences. We extracted DNA from goral feces, tissues, and blood samples, then amplified the AMEL (SE47/SE48 and SE47/SE53 primer pairs) and DDX3 genes for sex identification, comparing the goral DDX3X and DDX3Y target sequences to those in cattle. Our results indicated that the tissue- and blood sample-derived AMEL amplicons showed an unspecific band pattern containing the sex-specific band in the case of both primer pairs we used, whereas the DDX3 amplicon showed only the sex-specific band. In the case of the feces samples, only the sex-specific band was amplified using both the AMEL and DDX3 primer pairs. However, we found that the DDX3 amplicon exhibited a clearer band pattern than the AMEL amplicon. Then, we compared the DDX3X and DDX3Y target sequences between cattle and gorals. We found 5 and 8 nucleotide differences in the DDX3X and DDX3Y sequences, respectively. In conclusion, the DDX3 gene-related sex identification of the long-tailed goral appears to be more efficient and precise than the AMEL gene-related approach. This method could be used for the sex identification of the members of the Bovidae family.

An unidentified moth was captured in sex pheromone traps of the oriental fruit moth, Grapholita molesta, especially at spring season in apple orchards and their vicinity. Though the captured males were similar in appearance to G. molesta males, they were easily distinguished by a marked difference in body size. Their occurrence pattern was also similar to that of overwintering G. molesta population from April to May, at which more males were captured in the pheromone traps installed in the vicinity of apple orchards than within apple orchards. After May, they were no longer captured in the pheromone traps. To investigate any larval damage due to this unidentified moth, molecular markers needed to be developed. Four PCR-RFLP markers originated from cytochrome b region of mitochondrial DNA could distinguish this unidentified moth from G. molesta.

수정란의 성 판별은 유전적으로 우수한 유전형질을 보유하고 있는 소의 수정란을 성 판별하므로서 희망하는 성의 송아지를 생산할 수 있으며, 부가가치가 높은 수정란을 확보할 수 있는 기술이다. 수정란의 손상을 최소화하면서 할구를 biopsy하는 기술을 개발하고, 간단하고 빠른 시간에 성 판별이 가능한 Loop-mediated isothermal amplification방법으로 수정란을 성 판별을 실시한 결과는 다음과 같다. 1. 한우 체내 수정란의 성비는 암

본 연구에서는 한우 태아의 시기별로 35일령, 50일령, 70일령 및 90일령의 fetal fibroblast cell line을 생산하였고, bovine-specific primer와 Y chromosome-specific primer를 이용하여 PCR에 의해 성을 판별하여 각각 암수 2 line의 한우 fetal fibroblast cell line을 확립하였다. 이들 cell line을 계대배양하여 passage number가 10 이상에서 염색체 분석을 실시하였는데 모두에서 80%이상의 세포가 60개의 정상 염색체수의 나타내어 계대배양이 karyotype에 영향을 미치지 않는 것으로 나타났다. Serum starvation과 confluent 배양 방법을 이용하여 Go 상태로 유도되었는지 확인하기 위해 PCNA antibody를 이용하여 Western blotting 분석을 실시하였는데 PCNA 발현이 현저히 감소되는 것을 확인할 수 있었고, 다시 정상 medium으로 환원시켰을 때 세포분열이 재개되어 Go상태로 유도되었음을 확인할 수 있었다. 또한 serum stravation 방법이 conflent한 배양방법보다 PCNA 발현양이 적은 것으로 나타나 좀더 효율적인 Go 상태 세포 주기 조절방법으로 판명되었다.

This study was carried out to determine the sex of genomic and embryonic DNA using polymerase chain reaction(PCR). Bovine specific(216bp) and Y chromosome speicific DNA primers(l4lbp) were synthesized and tested for sexing. Bovine embryos used in this study were produced by in vitro fertilization. Few blastomeres for PCR were bisected by nicromanipulator and demi -embryos were cultured in TCM 199 medium containing 0.1% of solcoseryl. The results obtained were as follows; 1. Average optical density of genomic DNA extracted from blood of Hanwoo was 1.79 0.14. 2. 2. The ratio of the demi-embryos developed to blastocyst was 62.1 and 81.9% in morula and blastocyst, respectively. 3. When DNA of 2~4, 5~10 and more than 11 blastomeres was amplified with Y chromosome specific DNA primer by PCR, appreance rate of Y specific DNA band was 16.7, 46.2 and 40.0%, respectively. At least 5 to 10 blastomeres were required to determine the sex of embryos. 4. The rate of demi-embryos developed to blastocyst was 73.3% in TCM 199 medium supplemented with 0.1% solcoceryl. but 55.6% in control.