• International Journal of Industrial Entomology and Biomaterials
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• 제34권1호
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• pp.11-15
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• 2017

We constructed the fibroin H-chain expression system to produce enhanced cyan fluorescent proteins (ECFP) in transgenic silkworm cocoon. Fluorescent cocoon could be made by fusing ECFP cDNA to the heavy chain gene and injecting it into a silkworm. The ECFP fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, was designed to be secreted into the lumen of the posterior silk glands. The expression of the ECFP/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworms. The EGFP fluorescence became visible in the ocelli and in the central and peripheral nervous system on the seventh day of embryonic development. A mixture of the donor and helper vector was micro-injected into 1,020 Kumokjam, bivoltin silkworm eggs. We obtained 6 broods. The cocoon was displayed strong blue fluorescence, proving that the fusion protein was present in the cocoon. Accordingly, we suggest that the ECFP fluorescence silk will enable the production of novel biomaterial based on the transgenic silk.

• International journal of advanced smart convergence
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• 제12권4호
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• pp.434-442
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• 2023

The relationship between acute kidney injury (AKI) prediction and nephrotoxic drugs, or drugs that adversely affect kidney function, is one that has yet to be explored in the critical care setting. One contributing factor to this gap in research is the limited investigation of drug modalities in the intensive care unit (ICU) context, due to the challenges of processing prescription data into the corresponding drug representations and a lack in the comprehensive understanding of these drug representations. This study addresses this gap by proposing a novel approach that leverages patient prescription data as a modality to improve existing models for AKI prediction. We base our research on Electronic Health Record (EHR) data, extracting the relevant patient prescription information and converting it into the selected drug representation for our research, the extended-connectivity fingerprint (ECFP). Furthermore, we adopt a unique multimodal approach, developing machine learning models and 1D Convolutional Neural Networks (CNN) applied to clinical drug representations, establishing a procedure which has not been used by any previous studies predicting AKI. The findings showcase a notable improvement in AKI prediction through the integration of drug embeddings and other patient cohort features. By using drug features represented as ECFP molecular fingerprints along with common cohort features such as demographics and lab test values, we achieved a considerable improvement in model performance for the AKI prediction task over the baseline model which does not include the drug representations as features, indicating that our distinct approach enhances existing baseline techniques and highlights the relevance of drug data in predicting AKI in the ICU setting.

• Molecules and Cells
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• 제40권11호
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• pp.828-836
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• 2017

Eukaryotic cells consist of a complex network of thousands of proteins present in different organelles where organelle-specific cellular processes occur. Identification of the subcellular localization of a protein is important for understanding its potential biochemical functions. In the post-genomic era, localization of unknown proteins is achieved using multiple tools including a fluorescent-tagged protein approach. Several fluorescent-tagged protein organelle markers have been introduced into dicot plants, but its use is still limited in monocot plants. Here, we generated a set of multicolored organelle markers (fluorescent-tagged proteins) based on well-established targeting sequences. We used a series of pGWBs binary vectors to ameliorate localization and co-localization experiments using monocot plants. We constructed different fluorescent-tagged markers to visualize rice cell organelles, i.e., nucleus, plastids, mitochondria, peroxisomes, golgi body, endoplasmic reticulum, plasma membrane, and tonoplast, with four different fluorescent proteins (FPs) (G3GFP, mRFP, YFP, and CFP). Visualization of FP-tagged markers in their respective compartments has been reported for dicot and monocot plants. The comparative localization of the nucleus marker with a nucleus localizing sequence, and the similar, characteristic morphology of mCherry-tagged Arabidopsis organelle markers and our generated organelle markers in onion cells, provide further evidence for the correct subcellular localization of the Oryza sativa (rice) organelle marker. The set of eight different rice organelle markers with four different FPs provides a valuable resource for determining the subcellular localization of newly identified proteins, conducting co-localization assays, and generating stable transgenic localization in monocot plants.

• Asian-Australasian Journal of Animal Sciences
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• 제22권4호
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• pp.492-499
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• 2009

A Silkie Bantam embryo fibroblast line (named SBF59 line) was successfully established by using direct explant culture and cryopreservation techniques. Cell morphology, viability, dynamic growth and contamination were tested and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analyzed. Four kinds of fluorescent protein extrogenes, including $pEGFP-N_3$, $pECFP-N_1$, $pEYFP-N_1$ and $pDsRed1-N_1$ were transfected into the cells. The results showed that the cells were healthy and possessed a fibrous structure without a change in morphology. The average viability of the cells was 96% before freezing and 90.5% after thawing. The growth curve appeared as typical "S" shape and the cell growth passed through a detention phase, a logarithmic phase and a platform phase; the estimated population doubling time (PDT) was 38.5 h; assays for the presence of bacteria, fungi, viruses and mycoplasmas were negative; the cell line showed no cross contamination when assessed by isoenzyme analysis; the chromosome number was 2n = 78 on more than 88% of occasions; four kinds of fluorescent protein extro-genes appeared to be expressed effectively with a high transfection efficiency between 18.3% and 42.3%. The cell line met the required quality control standard. It not only preserves the genetic resources of the important Silkie Bantam at the cellular level but also provides valuable materials for genomic, post-genomic, somatic cell cloning research and other applications.

• 한국가축번식학회지
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• 제26권3호
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• pp.205-214
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• 2002

현재까지 외래 유전자를 도입하여 형질전환 동물을 생산하는 방법이 다방면으로 연구되어 왔다. 그 중에서 본 연구에서는 정자를 EGFP 유전자와 공배양한 후 이를 난모 세포내에 미세 주입한 다음, 수정란의 발달과 ECFP 유전자의 발현을 조사하였다. 즉, 동결후 융해나 Triton X-100 처리 등으로 세포막을 파괴하여, 이들 정자를 EGFP 유전자와 다분간 공배양함으로써 정자와 EGFP 유전자와의 결합을 유도하였다. 정자나 정자두부의 미세주입에 의해 수정된 난자는 0.3%의 BSA가 첨가된 CR1aa 배양액에서 배양하였으며, EGFP 유전자의 발현은 형광현미경 하에서 관찰하였다. 통결 후 융해로 처리된 정자와 Triton X-100 처리 한 정자를 미세주입한 결과 난할율은 85.7과 80.1%였고, 배반포 발생율은 32.4 과 35.0%로서 유의차가 없었다. 동결 후 융해와 Triton X-100 으로 처리된 정자를 각각 미세주입한 수정란의 EGFP 유전자 발현율은 각각 19.1과 13.9%로서 전자가 유의하게 높았다. 또 정자 배양액에 첨가된 EGFP유전자의 농도가 54 ng/${\mu}\ell$일 때 EGFP 발현율은 15.4% 로서, 27 ng/${\mu}\ell$일 때의 9.0%와 63.5 ng/${\mu}\ell$일 때의 5.1% 보다 유의하게 높았다. 발현율을 높히기 위한 방법중 하나로써 electric shock의 방법을 이용해 보았으나 기존의 공배양 방법으로 얻은 최고 발현율인 19.1%에 못 미치는 2%를 보였다. EGFP 유전자가 발현된 수정란의 배반포 발생율은 0%로서 비발현 수정란의 29.5%보다 유의하게 낮았으며, EGFP 유전자의 발현은 mosaicism 형태를 보였다. 본 연구에서는 비록 낮은 외래 유전자 도입율을 보이기는 하나 (19.0%), 정자를 매개로 한 형질전환 동물의 생산은 그 방법이 간단하고 비용이 적게 든다는 장점이 있다. 기존 보고들의 효율성을 재고하여 볼 때, 난자내 정자 직접 주입술에 의한 형질전환 동물 생산의 연구는 향후 밝은 전망을 시사하고 있다.