• Title/Summary/Keyword: $BaCO_3$

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Protoplast Fusion of Nicotiana glauca and Solanum tuberosum Using Selectable Marker Genes (표식유전자를 이용한 담배와 감자의 원형질체 융합)

  • Park, Tae-Eun;Chung, Hae-Joun
    • The Journal of Natural Sciences
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    • v.4
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    • pp.103-142
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    • 1991
  • These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

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Effect of an Acid pH Shock on Physiological Changes of Chlamydomonas acidophila (Chlorophyta), UTCC 122 (pH 충격에 의한 Chlamydomonas acidophila (Chlorophyta), UTCC 122의 생리적 변화에 관한 연구)

  • Lee, Kyung;Ki, Jang-Seu;Kim, Say-Wa;Han, Myung-Soo;Choi, Young-Kil;Yoo, Kwang-Il
    • Korean Journal of Ecology and Environment
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    • v.35 no.3 s.99
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    • pp.145-151
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    • 2002
  • The effect of low pH on physiological changes was studied with the acidophilic green alga, Clamydomonas acidophila, UTCC 122. The growthrates (${\mu}$) were identical, $0.5{\sim}0.7\;day^{-1}$, at pH 3.7${\sim}$6.7 and no significantly different (ANOVA, p =0.134), showing cell volume reduced gradually as they were growing, whereas that at pH 2.7 was falling to zero and cell volume increased dramatically. Chlorophyll a concentration of the cultures incubated for one day was $191{\sim}255\;pg\;cell^{-1}$, after then it declined from $60{\sim}103\;pg\;cell^{-1}$ at pH 3.7${\sim}$6.7 except $210\;pg\;cell^{-1}$ at pH 2.7, which was directly related with cell volume. External carbonic anhydrase (CA) activity was varied from1.1 to$3.7{\times}10^{-4}\;E.U.\;mm^{-2}$, showing the gradualincrease during culture, except at 2.7 and pH 5.7. However there was not found any relationship among the pH gradient cultures. CA molecular mass of C. acidophila was 29 kBa, and concentration of that was identical in all cultures. The proteins of 41 kDa and 63 were not or very faintly expressed in low pH cultures, in contrast that of 17 kDa more expressed. In this work, we found that C. acidophila could live optimally within a wide range of acidic pH, and 17 kDa of unidentified protein might be concerned with tolerating in low acid environment.

Changes of Proteolytic Activity and Amino Acid Composition of the Tissue Extract from Sea Cucumber Entrails during Fermentation with Salt (해삼내장(內臟)젓갈 숙성중(熟成中) 단백질분해효소(蛋白質分解酵素)의 활성(活性)과 아미노산(酸) 조성(組成)의 변화(變化))

  • Lee, Gi Chan;Cho, Deuk Moon;Byun, Dae Seok;Joo, Hyen Kyu;Pyeun, Jae Hyeung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.12 no.4
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    • pp.342-349
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    • 1983
  • This study was undertaken to ascertain food and nutritional evaluating data on the processing of fermented sea cucumber (Stichopus japonicus) entrails. In the experiment, the crude proteolytic enzyme from the entrails tissue of raw and fermented sea cucumber during the days of ripening was extracted. The optimal activity condition for the crude enzyme and the compositional changes of amino acid of the protein and free amino acid in the raw and fermented sample were also investigated. 1. Less than three kinds of proteolytic enzymes that each enzyme has optimal activity condition at pH 3.1 $50^{\circ}C$(A-enzyme), pH 5.7 $50^{\circ}C$(B-enzyme) and pH 7.7 $45^{\circ}C$(C-enzyme), respectively were believed to be exist in the entrails tissue of sea cucumber. 2. A-enzyme and C-enzyme were strongly inhibited with the increase of the salt concentration, and B-enzyme was activated at the 1% salt concentration and was inhibited above the 5% salt concentration. 3. The result of the effect of several salt ions on the proteolytic activity showed that A-enzyme was slightly inhibited in the presence of all salt ions added, B-enzyme was activated in the presence of the all salt ions except $Cu^{2+}$ and C-enzyme was activated in the presence of $Ca^{2+}$ and $Mn^{2+}$, and inhibited by $Cu^{2+}$, $Co^{2+}$ and $Mg^{2+}$. 4. When the effects of the ripening days on the proteolytic activity of the crude enzymes were analysed, the activity of the A-enzyme was slightly weakened with the lapse of the fermentation days, whereas the B-enzyme was not influenced by the fermentation days. 5. In the analysis of amino acid composition of the protein of the samples, the 8 days fermented sea cucumber entrails showed the diminution of all kinds of amino acid. Apparently diminished amino acids were arginine, alanine, glutamic acid, glycine, serine, valine, threonine and lysine etc., and methionine, histidine and isoleucine were slightly decreased. 6. In the analysis of free amino acid composition of the 8 days fermented sample, glutamic acid, aspartic acid, leucine and lysine were rich, while histidine, methionine, proline and tyrosine were poor. The most of free amino acids were increased during the fermentation procedure and especially in lysine, histidine, threonine, glutamic acid, methionine, valine and leucine.

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Results of Radiotherapy for the Uterine Cervical Cancer (자궁경부암의 방사선치료성적)

  • Kim, Chul-Yong;Choi, Myung-Sun;Suh, Won-Hyuck
    • Radiation Oncology Journal
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    • v.6 no.1
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    • pp.63-73
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    • 1988
  • One hundred fifty-four patients with the carcinoma of the uterine cervix were studied retrospectively to assess the result and impact of treatment at Department of Radiation Oncology, Korea University, Hae-Wha Hospital from Feb 1981 through Dec. 1986. Prior to radiotherapy, the patients were evaluated and staged by recommendation of FIGO including physical examination, pelvic examination, cystoscopy, rectosigmoidoscopy, chest X-ray, IVP. Ba enema. Also, an additional pelvic CT scan was obtained for some of the patients. The patients were treated by radiotherapy alone or adjuvant postoperative irradiation; in case of radiation therapy only, whole pelvic irradiation was given with Co-60 teletherapy unit via AP and PA parallel opposing fields or 4-oblique fields, 180 cGy per day, 5 days per week and intracavitary insertion was performed. In satges Ia, Ib, and IIa with small primary lesion, external irradiation was initially given to pelvis up to $2,000\~3,000\;cGy/2frac{1}{2}\;-3frac{1}{2}$ weeks and then intracavitary insertion was performed using Fletcher-Mini-Declos Applicator with cesium-137 cources and followed by external irradiation of $1,000\~2,000\;cGy/1frac{1}{2}\;-2frac{1}{2}$weeks via AP and PA parallel opposing fields with midline shield to spare of bladder and rectum. However, if the primary lesion is large, external irradiation was given without midline shield. More than stages IIb, the patients were treated by external beam irradiation up to 5,400cGy/30f for 6 weeks via 4-oblique portals and at the dose of 5,040cGy/28f the field was cut 5cm from the top margin for spare of small bowel, and followed by intracavitary irradiation, If there was residual tumor an additional dose of $900\~l,200cGy/5\~7f$ was given to parametrium and/or residual tumor area. Total dose of radiation to A and B-point were as follows; A-point; In early stages, Ia, Ib, IIa; $8,000\~9,000$ B-point $5,000\~6,000 cGy$ A-point; In advanced stages IIb, IIIa, IIIb; $9,000\~10,000$ B-point $60,000\~7,000cGy$ The results were obtained and as fellows; 1 The patients distribution according to FIGO staging system were stage Ia 6, Ib 27, IIa 28, IIb 54, IIIa 12, IIIb 18, and stage IVa 9. 2. Value of CT scan were demonstration of cervix tumor mass, parametrial and pelvic side wall tumor spread, pelvic and inguinal lymph nodes metastases, and hydronephrosis. Three dimensional quantitative demonstration of tumor volume is also important in planning radiation therapy. Another advantage of CT scan was detection of recurrent tumor after radiation or surgery. 3. Local control rate of tumor according to the size was $91.3\%$ for less than 5cm in size and $44.6\%$ in tumor over 5cm (p<0.0068). 4. Thirty out of 50 recurrent sites has locoregional failures and 17 cases has distant metastases. And the para-aortic lymph nodes were the most common site for distant metastases. 5. The most common complication was temporal rectal bleeding which was controlled most by conservative management. However, 4 patients required for endoscopic cauterization. 6. The 5-year survival rates showed; stage la and Ib $95\%,\;stage\;IIa\;81\%\;stage\;lIb\;67\%,\;stage\;IIIa\;37.7\%,\;stage\;IIIb\;23\%$ and 3-year survival rate of stage IVa showed $11.6%$, retrospectively.

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Reproductive Cycle of the Echiuroid Worm Urechis unicinctus(von Drasche) in Southern Korea (한국산 개불, Urechis unicinctus (von Drasche)의 생식주기)

  • 최상덕;김호진;이원교;곽은주;윤호섭;라성주;이인곤
    • Journal of Aquaculture
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    • v.13 no.2
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    • pp.169-174
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    • 2000
  • Reproductive cycle of U. unicinctus was studied from September 1998 to August 1999, using gonadosomatic index (CSI) as an indicator. In November, the CSI values were maximum for male (6.2) and female (7.0), respectively; the values were lowest for them (1.0 and 0.5) during the successive february. Subsequently, they rapidly increased and attained peak by March-April. The values decreased again in both sexes and remained unchanged until August. The index increased in October to attain the peak by November. The CSI values clearly indicated that there are two spawning events in a year, namely the first one during April-May and the second one in December. Reproductive cycle was classified into the following successive stages: in female, multipication (January~February, June ~Setember), maturation (March~April, November), spent (May and December), degeneration and resting (June and January), and in male, multiplication January ~ february, June ~September), maturation (March~April, October~November), spent (May and December) and degeneration and resting (January and June). Histological observations revealed that oocytes in the ovary matured simultaneously in November and March. At the same time, the envelopes of matured testis became thinner than those in the early stage.

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Characterization of Chitin Deacetylase Produced from Mucor rouxii (Mucor rouxii가 생산한 Chitin deacetylase의 특성)

  • SOHN Heung-Sik;PARK Seong-Min;SON Byung-Yil;CHOI Hyeon-Mee;LEE Keun-Tai
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.2
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    • pp.121-126
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    • 1999
  • In order to degrade chitin by enzymatic hydrolysis, it is required from screening highly active deacetylase. To this end, we examined three fungal strains and it turned out that Mucor rouxii produced highly active deacetylase, this enzyme exhibited the highest enzymatic activity against colloidal chitin. The conditions for growing Mucor rouxii are as follows; the effective carbon source, nitrogen source, adequate initial pH, temperature and incubation time were $2\%$ glucose, $1.33\%$ yeast extract, $0.66\%$ pepton, 4.5, $25{\pm}2^{\circ}C$ and 48hr, respectively. The optimum pH and temperature for purified enzyme activity were 5.5 and $40^{\circ}C$, respectively. The purified enzyme was stable at pH ranging from 4.5 to 5.5. However, the enzyme activity was decreased to less than $50\%$ at pH blow 45 and above 7.5. At temperatures above $50^{\circ}C$, the enzyme activity was decreased remarkably. The enzyme was inhibited by LiC1, $HgCl_2$, and $BaCl_2$, but stimulated by $CaCl_2$ and $ZnC1_2$, The activity of purified enzyme was increased by L-cysteine and 2-mercaptoethanol, while decreased by O-phenanthroline, p-CMB, EDTA, and iodoacetate. The $K_m$ and the $V_{max}$ value of purified enzyme were $1.2\%$ and 59.5 U/mg, respectively. The deacetylation activity of purified enzyme was not detected at optimal reaction condition when chitin particle suspension was used.

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