• 원예과학기술지
• /
• 제29권2호
• /
• pp.123-129
• /
• 2011

To identify unintended vertical gene-transfer rates from the developed transgenic plants, rapid and unequivocal techniques are needed to identify event-specific markers based on flanking sequences around the transgene and to distinguish zygosity such as homo- and hetero-zygosity. To facilitate evaluation of zygosity, a polymerase chain reaction technique was used to analyze a transgenic pepper line B20 (homozygote), P915 wild type (null zygote), and their F1 hybrids, which were used as transgene contaminated plants. First, we sequenced the 3'-flanking region of the T-DNA (1,277 bp) in the transgenic pepper event B20. Based on sequence information for the 3'- and 5'-flanking region of T-DNA provided in a previous study, a primer pair was designed to amplify full length T-DNA in B20. We successfully amplified the full length T-DNA containing 986 bp from the flanking regions of B20. In addition, a 1,040 bp PCR product, which was where the T-DNA was inserted, was amplified from P915. Finally, both full length T-DNA and the 1,040 bp fragment were simultaneously amplified in the F1 hybrids; P915 ${\times}$ B20, Pungchon ${\times}$ B20, Gumtap ${\times}$ B20. In the present study, we were able to identify zygosity among homozygous transgenic event B20, its wild type P915, and hemizygous F1 hybrids. Therefore, this novel zygosity identification technique, which is based on PCR, can be effectively used to examine gene flow for transgenic pepper event B20.

• Advances in pediatric surgery
• /
• 제15권2호
• /
• pp.157-160
• /
• 2009

Although the incidence of esophageal atresia (EA) is higher in twins than in singletons by two to three times, EA usually affects only one member of twins. We report one pair of twins concordant for EA. A 31-year-old healthy woman bore monozygotic female twins at 36 weeks of gestation. They weighed 2,216 and 2,480 g, respectively. They had EA with distal tracheoesophageal fistula and underwent primary esophageal anastomosis on the birth day and the $2^{nd}$ day of life, respectively. Twin A also had suspicious antral obstruction and pyloroplasty was done simultaneously with esophageal repair. She needed antral web excision for continued gastric stasis one month after $1^{st}$ operation and three balloon dilatations of the esophagus. Twin B recovered uneventfully.

• 한국동물생명공학회지
• /
• 제39권2호
• /
• pp.81-87
• /
• 2024

Background: Platelet-derived growth factor receptor alpha (PDGFRα) is essential for various biological processes, including fetal Leydig cell differentiation. The PDGFRα^EGFP mouse model, which expresses an eGFP fusion gene under the native Pdgfrα promoter, serves as a valuable resource for exploring PDGFRα's expression and function in vivo. This study investigates PDGFRα expression in adult testicular cells using PDGFRα^EGFP mouse model. Methods: Genotyping PCR and gel electrophoresis were used to confirm the zygosity of PDGFRα^EGFP mice. Histological examination and fluorescence imaging were used to identify PDGFRα expression within testicular tissue. Immunohistochemical analysis assessed the co-expression of PDGFRα with c-Kit, ANO-1, and TASK-1 in testicular cells. Results: Genotyping confirmed the heterozygous status of the mice, which is crucial for studies due to the embryonic lethal phenotype observed in homozygotes. Histological and fluorescence imaging revealed that PDGFRα⁺ cells were primarily located in the interstitial spaces of the testis, specifically within Leydig cells and peritubular myoid cells (PMCs). Immunohistochemical results showed PDGFRα co-localization with c-Kit and ANO-1 in Leydig cells and a complete co-localization with TASK-1 in both Leydig cells and PMCs. Conclusions: The findings demonstrate specific expression of PDGFRα in Leydig cells and PMCs in adult testicular tissue. The co-expression of PDGFRα with c-Kit, ANO-1, and TASK-1 suggests complex regulatory mechanisms, possibly influencing testicular function and broader physiological processes.

• Journal of Genetic Medicine
• /
• 제9권2호
• /
• pp.73-77
• /
• 2012

Purpose: Fragile X carrier detection before or at early pregnancy through a wide screening program may not only confer a risk of having offspring with Fragile X syndrome (FXS), but may also confer a risk for Fragile X-associated primary ovarian insufficiency and Fragile X-associated tremor/ataxia syndrome. However, prior to the implementation of such a program, the carrier prevalence in a population and the availability of effective screening test should be evaluated. The aim of our study was to determine the prevalence of premutation carriers and to evaluate the feasibility of screening test. Materials and Methods: The blood samples were obtained from 8,641 pregnant women with no family history of mental retardation. We performed a three-primer CGG repeat primed (RP) PCR using the AmplideX$^{TM}$ FMR1 PCR kit (Asuragen, Inc. Austin, TX, USA). Samples showing full mutation alleles were reflexed to Southern blot analysis for methylation status and sizing. Results: Among the 8,641 women, we found 8 premutation carriers (1:1,090, 0.09%) and 46 women with an intermediate allele (1:190, 0.53%). No woman was found to carry the fully mutated allele. All the detected alleles were within the CGG repeat range of 8-117. Among the 8,641 samples, 29 and 30 CGG repeats represent 66.6% of all cases. The CGG RP PCR method provides robust detection of expanded alleles and resolves allele zygosity, thus minimizing the number of samples that require Southern blot analysis. Conclusion: This is the first study that has focused on the prevalence of FXS premutation carriers and FMR1 allele distribution in normal pregnant women. These data have important implications for population-based fragile X carrier screening in Korea.