• Biotechnology and Bioprocess Engineering:BBE
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• 제10권4호
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• pp.353-356
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• 2005

The effect of silkworm hemolymph on the expression of recombinant protein in Escherichia coli was investigated. The addition of silkworm hemolymph to the culture medium in­creased the production of recombinant $\beta$-galactosidase in E. coli. The production was dependent on the concentration of the added silkworm hemolymph, which increased 2-, 5-, and 8-fold in media supplemented with 1, 3, and $5\%$ silkworm hemolymph, respectively. To identify the effective component, the silkworm hemolymph was fractionated by gel filtration column chromatography. A fraction, with a molecular weight of about 30 K was identified as the effective component.

• 한국동물학회지
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• 제12권2호
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• pp.60-66
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• 1969

배추흰나비(Pieris rapae Linne)의 變態에 따른 hemolymph protein 의 濃度變化와 酸素消費量 및 5齡의 呼吸基質에 對한 活性度를 各各 Biuret method 와 Warburg manometric method 로 測定하여 形態的 變化를 比較하였다. 1. Hemolymph protein 의 濃度는 終齡幼蟲에서 가장 높으며 變態에 따라 감소하여 용기말에서 다시 增加한다. 2. Endogenous respiration은 전용기에서 가장 높은 酸素消費量을 보이며 變態에 따라 감소하여 용기말에서 다시 增加한다. 3. Glucose 는 全變態期를 通하여 다른 呼吸期質보다도 가장 큰 活性을 보이며 전용과 용기말에서 현저하게 영향을 미친다. 4. Hemolymph protein 의 濃度變化와 酸素消費量은 全變態期를 通하여 밀접한 연관을 가지며 幼蟲器官의 解消와 成蟲器官의 新生에 따라 U字型을 나타낸다.

• 한국동물학회지
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• 제37권1호
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• pp.66-75
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• 1994

Juvenile hormone binding protein was identified in the hemolymph of fifth instar larvae and purified using column chromatography. Hemolymph was mixed with [3H] JH-III and electrophoresed on 691 NON-SDS gel, indicating that radioactivity peak appears at Rf value of 0.55. Gel filtration showed two radioactivity peaks equivalent to bound and free [3H]JH-III, respectively. JHBP was purified from hemolymph through gel filtration (Sephadex G-100), anion exchange chromatosraphv (DEAE Sepharose CL-6B), chromatofocusing chromatographv (PBE 94) and preparative electrophoresis.

• 한국잠사곤충학회지
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• 제30권1호
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• pp.25-32
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• 1988

누에 유충에 있어서 체액 JH 결합단자질(JHBP)의 생리적 역할을 알기 위하여, 체액 중의 JH 1과 결합하는 단자질을 polyacrylamide gel 전기영동으로 분리하고, 또 발육에 따른 JH 결합단자질의 활성 변화를 측정한 결과는 다음과 같다. 1. 누에 체액중에서 JH 1과 결합하는 단자질은 JH 결합지질단자질(JH-LP)과 Jh 결합단자질(JHBP) 등 JH결합단자질의 Rm은 0.81∼0.33이었고, JH 결합단자질의 Rm은 0.81∼0.84이었다. 두 단자질의 Rm은 성별, 발육시기에 관계없이 일정하였다. 2. 체액단자질의 함량은 발육의 진행에 따라 점차 증가하여 토사기에 가장 높았으며 전용기에 다소 감소하였고 암컷의 함량은 숫컷보다 높았다. 3. 체액 ml 당 JH 결합단자질의 활성은 영의 초기에 낮았고 토사기에 최대로 나타난 후 전용기에 다소 감소하였다. 4. 체액 ml 당 JH 결합단자질의 활성 변화는 체액단자질의 함량의 변화양상과 매우 유사하였다. 5. 체액단자질 mg 당 JH 결합단자질의 활성은 영의 초기에 높았고 5령 6일에서 전용기까지 낮은 수준을 유지하였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.62-65
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• 2002

We have previously shown that the addition of silkworm hemolymph to a culture medium increases the longevity of insect and mammalian cells by inhibiting apoptosis. This indicates that the component which inhibits apoptosis is contained in the silkworm hemolymph, The apoptosis-inhibiting component was isolated from silkwonn hemolymph and characterized in our previous study. A database search using the N-terminal amino acid sequence of this component as a template resulted in a 95% homology with a low molecular weight lipoprotein, the so called ’30K protein' of unknown function. In this study, the 30K protein gene was expressed in mammalian and insect cells to confirm the apoptosis-inhibiting effect. The overexpression of 30K protein in mammalian cell inhibited the staurosporin-induced apoptosis by the prevention of the activation of caspase 3. Using an Autographa californicanuclear polyhedrosis virus (AcNPV) system, the 30K protein was overexpressed also in insect cells. The expression of the 30K protein increased the longevity of baculovirus-infected insect cells by inhibiting apoptosis. These results suggest that the 30K protein is a novel anti-apoptotic protein.

• International Journal of Industrial Entomology and Biomaterials
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• 제8권1호
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• pp.117-121
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• 2004

Soluble proteins of silkworm hemolymph were investigated by means of isoelectric focusing (IEF). The protein spectra during ontogenesis of races and inter-races hybrids kept in Bulgaria was studied. A total of 51 protein bands in the hemolymph from fourth larval instar to imago were ascertained. Stage specific expression was established. The specific expression of some protein bands in the individual spectra manifest phenotype of gene determinate polymorphism (HP F, HP J, HP K, HP L, HP Q - in the zone with pH gradient 3.5-6.2 and HP K, HP L, HP N, HP P, HP T - in the zone with pH gradient 9.5 - 6.2). Breed specific expression was observed. On the basis of the obtained results, it was established that the investigated breeds are heterogeneous and the isoelectric focusing method is successful when specifying the inner-race and inter-race polymorphism in silkworm.

• Animal cells and systems
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• 제1권1호
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• pp.71-75
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• 1997

Male specific protein (MSP) was identified and purified from the hemolymph of Galleria melloneffa L. by electrophoresis and anion exchange chromatography. MSP has a native molecular weight of 55 kDa as determined by gel filtration chromatography and consists of a single unit with the apparent molecular weight of 27 kDa and has the pl of approximately 5.8. MSP is present in the hemolymph from day 8 pupae throughout the male adult. MSP was also found in pupal fat body, adult fat body, and adult testis.

• 생명과학회지
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• 제31권4호
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• pp.425-429
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• 2021

INS-1 췌장 β 세포에서 Allomyrina dichotoma 혈림프 처리에 의한 early growth response protein 1 (EGR1) 유전자 발현을 조사되었다. 이 연구에서 새로운 발견은 EGR1 유전자 발현을 A. dichotoma 혈림프의 용량 및 시간 의존적으로 상향 조절하는것과 혈림프와 병행한 저체온효과 또는 소포체(endoplasmic reticulum, ER) 스트레스에 의해서도 유전자 발현이 상승하였다. A. dichotoma 혈림프가 EGR1의 유전자발현을 상승 조절을 할 수 있기 때문에, EGR1 관련 질병 치료 및 예방의 실마리를 제공 할 수 있을 가능성을 시사한다.

• Animal cells and systems
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• 제3권2호
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• pp.173-180
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• 1999

A blue biliprotein, insecticyanin (INS), has been purified from the last instar larval hemolymph of Agrius convolvuli by ultracentrifugation, Sephadex G-100 gel permeation chromatography, and preparative electrophoresis. The molecular mass of INS was estimated to be 26 kDa and the N-terminal sequence of INS revealed high similarity to that of Manduca sexta. Results of Western blotting and autoradiography indicated that INS is synthesized by the epidermis and released into the hemolymph. In contrast to the INS reported in other insects, Agrius convolvuli INS contained a small amount of lipid, predominately consisting of triacylglycerol. Subcellular localization of INS was determined using protein-A gold particles linked to secondary antibodies (anti-rabbit Ig). INS was heavily accumulated in the cytoplasmic inclusion body (CIB). CIBs showed a variety of shapes from rod to globule and generally surrounded the nucleus. They were mostly located near the basement membrane and especially abundant in the intersegmental membrane.

• International Journal of Industrial Entomology and Biomaterials
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• 제5권2호
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• pp.189-194
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• 2002

Silkworm diseases are major constraint in silk cocoon production. Among silkworm diseases, white muscar-dine is highly contagious and most common in winter and rainy seasons. It is suggested that hemocytes involve in defense against invasion of Beauveria bassi-ana and systemic fungicide/chemicals prevent the proliferation of fungi in the hemolymph or preventing the growth of the fungi in the body cavity through enhancing the hemocyte mediated defense response. In the present study the influence of systemic fungicide on hematological changes in silkworms infected with Beauveria bassiana was reported. It is observed that the total hemocyte counts increased in the hemolymph up to 5 days post inoculation in systemic fungicide treated batches while in the inoculated control the increase was up to 3 days indicating the positive hemocyte mediated response in silkworm treated with systemic fungicide. After 2 days in the inoculated control as the multiplication and growth of mycelia increased, defense capacity of the silkworm was decreased. The biochemical changes were also observed in the hemolymph of silkworm infected with B. bassiana. In silkworm infected with Beauveria bassi-ana, the total protein content increased whereas total carbohydrate and total lipids decreased as the infection progresses. In the case of systemic fungicide treated batches the increase in total protein content was comparatively higher and decrease in total carbohydrate and lipids were comparatively lower than the inoculated control.