• 생명과학회지
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• 제7권1호
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• pp.49-58
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• 1997

Engralied 5.7kb upstream sequence와 E. colilacZ의 융합 유전자를 가진 P[en-lacZ] 인자를 jumpstart 기법을 이용하여, ryXho25 strain의 초파리 48A 염색체 위치로부터 새로운 위치로 삽입하였다. 총 3315의 유전적 교배를 통해서, P[en-lacZ] 가 다른 염색체 상으로 삽인된 113 계통을 얻었다. X-gal 염색으로 이들 113 계통의 3령기 유충 조직에서의 $\beta$-galactosidase 발현을 조사하였다. 도한 113 계통 중 7계통이 열성치사돌연변이인 것으로 동정되었다. 이들 7 계통 중 초기 배발생 과정에서 치사하는 것으로 조사된 #1119의 초기 배발생 과정에서의 ${\beta}$-galactosidase 발현과 핵의 이동 및 세포화 양상을 조사하였다. 본 연구에서 얻어진 P[en-lacZ] 삽입 돌연변이체들은 앞으로 Drosophila 발생에 관련된 유전자들의 구조와 기능을 연구하는데 활용될 수 있을 것이다.

• KSBB Journal
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• 제10권1호
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• pp.38-45
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• 1995

Aspergillus oryzae에서 A. nidulans의 glyceral d dehyde-3-phosphate dehydrogenase (gpdA)와 trpC, prmoter의 발현 능력 을 E. coli lacZ gene fusion을 이용하여 비교.분석하였다. A. oryzae 내에서 발현된 E. coli $\beta$galactosidase의 specific activIty를 조사하여 본 결과, gpdA promoter를 가지는 transformant들 에서는 2,000unit/ug of protem 정도의 activity를 보이는 반면, trpC, promater를 가지고 있는 transformant들에서는 10.5~52.3unit/ug of protein 정도의 activity를 보였다. 이 결과로부터 A. oryzae 내에서 A. nidulans의 gpdA promoter가 trpC, promoter에 비해 70 배 정도 더 강한 발현 능력을 보이고 있음을 알 수 있다. Western blot 분석에서도 gpdA promoter를 가지고 있는 transf ormant에서 더 많은 E. coli $\beta$-galactosidase가 발현된 것 을 보여 주고 있다. 또한 southern blot 분석에서는 이러한 강한 발현이 transform된 plasmid의 copy number와 상관 없음을 보여주고 있다.

• Journal of Plant Biology
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• 제27권2호
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• pp.43-50
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• 1984

Effects of IAA on the elongation and cell wall hlysocidase activities were investigated in excised rape (Brassica napus L. cv. Yongdang) hypocotyl segments. IAA promoted the elongation of rape hypocotyl segments. In rape hypocotyls, the first 10-mm segments from the hook exhibited maximal elongation and the capacity of elongation was gradually decreased with increasing distance of each 10-mm from the hook. A good correlation has been obtained between the magnitude of endogenous growth and the activities of $\alpha$, $\beta$-glucosidase and $\alpha$, $\beta$-galactosidase. However, exogenous application of IAA did not seem to enhance the tissue with IAA resulted in acidification of the incubation medium. From these data, we can conclude that IAA seems to enhance elongation of the tissue segments, at least in part, by releasing hydrogen ion into cell wall, some of which may participate in the cell wall extension process, but does not seem to trigger the activation of $\alpha$, $\beta$-glucosidase and $\alpha$, $\beta$-galactosidase.

• 한국수산과학회지
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• 제31권5호
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• pp.637-644
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• 1998

우리나라 근해에 많이 분포하고 있는 말똥성게 (sea urchin, Hemicentrotus pulcherrimus)의 내장으로부터 Triton X-100을 이용하여 $\beta$-galactosidase를 추출하고, $40\~80\%$ (w/v) $(NH_4)_2SO_4$, DEAE-Sephadex A-25 및 CM-Cellulose 이온교환 크로마토그래피, Con A-Sepha-rose 4B 친화성 크로마토그래피를 사용하여 분리, 정제하여 그 생화학적 특성을 조사한 결과는 다음과 같다. 정제된 $\beta$-galactosidase는 단일의 단백질로 이루어진 효소로 판명되었고, 효소의 정제도는 조효소에 비해 384.6배 증가하였고, 수율은 $1.26\%$이었다. 정제효소의 최적 pH와 온도는 각각 3.0 및 $50^{\circ}C$ 이었다. 효소의 활성은 $Ba^{2+}$와 같은 금속이온에 의해 촉진되었고, $Hg^{2+},\;Sn^{2+}$ 및 DFP에 의해 현저하게 저하되었으며, 당인 galactose 와 lactose에 의해 저하되어 기질 저해 효과가 나타남을 알 수 있었다. 효소의 분자량은 SDS-PAG 전기이동과 Sephadex G-150 겔여과를 실시한 결과 97 kDa로 나타났다. 합성기질인 PNPG를 사용하여 효소의 속도론적 상수를 측정한 결과 $K_m$은 15.0mM, $V_{max}$는 $\mu$mole/min$\cdot$mg$\cdot$protein으로 나타났다.

• Molecules and Cells
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• 제20권1호
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• pp.43-50
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• 2005

Glutaredoxin (Grx) is a small, heat-stable redox protein acting as a multi-functional glutathione (GSH)-dependent disulfide oxidoreductase. We have cloned the monothiol Grx5 gene from the genomic DNA of the fission yeast Schizosaccharomyces pombe. It has 1,904 bp, with one intron, and encodes a putative protein of 146 amino acids with a molecular mass of 16.5 kDa. Recombinant Grx5 produced functional Grx in S. pombe cells. NO-generating sodium nitroprusside (SNP, 1.0 and 2.0 mM) and potassium chloride (KCl, 0.2 and 0.5 M) increased the synthesis of ${\beta}$-galactosidase from a Grx5-lacZ fusion gene, and transcription of Grx5 was also enhanced by SNP and KCl. Synthesis of ${\beta}$-galactosidase from the Grx5-lacZ fusion was lower in Pap1-negative TP108-3C cells than in wild type KP1 cells, and when Pap1 was overproduced in KP1 cells, the level of ${\beta}$-galactosidase increased. We also found that Pap1 is involved in the induction of Grx5 by SNP and KCl. S. pombe Grx5 may play a crucial role in responses to nitrosative and osmotic stresses.

• Food Science and Biotechnology
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• 제17권3호
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• pp.514-518
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• 2008

After overproduction of a recombinant $\beta$-galactosidase of Bifidobacterium infantis in Pichia pastoris, a synthesis of galacto-oligosaccharides (GOS) from 36% lactose using the enzyme (170.74 U/mg) was investigated. The transgalactosylation ratio reached up to 25.2% with 83.1% conversion of initial lactose and the maximum yield of GOS was 40.6%. The GOS syrup was composed of a 13.43% galacto-oligosaccharides, 5.06% lactose, and 8.76% monosaccharides. The prebiotic effect of GOS on the growth of bifidobacteria and lactobacilli strains was investigated in vitro. The maximum growth rate of Bifidobacterium breve and Lactobacillus acidophillus in GOS syrup (5%, v/v) media were 0.49 and 0.96/hr that are higher than those in 1%(w/v) galactose and 1%(w/v) lactose containing media. However, there was no significant difference between the specific growth rates of L. acidophillus in 1%(w/v) glucose and 5%(v/v) GOS syrup. Our data showed that GOS definitely promoted the growth of B. breve ATCC $15700^T$ and L. acidophilus ATCC 33323.

• 한국미생물·생명공학회지
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• 제24권4호
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• pp.493-499
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• 1996

Lactan gum produced by Rahnella aquatilis is a high viscous, anionic polysaccharide and has shear thinning behaviour. Lactan gum yield and cencentration was greater on disaccharide such as lactose and sucrose than on monosaccharides such as glucose and galactose. When initial carbon source concentration was 45g/l of sucrose of lactose, the microorgnisms produced 28 g/l and 27 g/l of lactan, respectively with a yield more than 60%. $\beta$-Galactosidase, hydrolyzing lactose into galactose and glucose, was induced by lactose or galactose. When initial corbon source was 45 g/l of mixed carbon I (glucose:galactose=1:1), lactan gum concentaration was higher than that from 45 g/l of monosaccharide (glucose pf galactose) but was similar to the result from 45 g/l of lactose. Therefore, lactose hydrolysis reaction by $\beta$-galactosidase does not seem to be a rate determining step in lactan gum biosynthesis. When initial carbon source was 45 g/l of mixed carbon II (glucose:fructose=1:1). total carbon source consumption rate was slower than that from sucrose, but glucose consumption rate was faster than that from fructose.

• 한국잠사곤충학회지
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• 제39권2호
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• pp.180-185
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• 1997

To develope the novel baculovirus transfer vector, the p10 gene was cloned from the Bombyx mori nuclear polygedrosis virus (BmNPV) vB2 strain isolated from the B. mori larvae of sericultural farms. The novel transfer vector was constructed by using the p10 gene of BmNPV vB2 strain was 210 bp. The TAAG sequence at the -71 bp of upstream from translation initiator ATG and two polyadenylation signal site at the downstream from terminator TAA were also detected in the p10 gene. The 5' and 3' flanking region of the p10 gene amplified by PCR was cloned into pBluescriptII SK(+) and then transfer vector pBm10 was construceted. The 7.9 kb pBm10 was analysed by restriction enzymes and the map was confirmed. In order to determine the expression of foreign gene of pBm10, $\beta$-galactosidase gene was inserted in the SmaI site of foreign gene cloning site of pBm10. The pBm10 containing $\beta$-galactosidase gene was cotranfected wth genomic DNA of BmNPV vB2 into BmN-4 cells. The recombinant baculovirus expressing $\beta$-galactosidase was also produced polygedra in the infected cells. The results indicated that pBm10 is functional, suggesting that in the baculovirus expression vector system, the recombinant virus produced by pBm10 was effective by oral infection for the producing recombinant proteins in in vivo expression.

• 한국잠사곤충학회지
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• 제37권2호
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• pp.181-185
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• 1995

국내 분리주의 BmNPV를 이용한 전이벡터 pBm-KSK1에 외래 유전자로서 E. coli $\beta$-galactosidase 유전자를 클로닝하여 제작한 재조합 바이러스 BmK1-lacZ의 발현 증대를 위해 재조합 바이러스 감염세포주에 누에 체액의 첨가에 따른 발현효율을 조사하였다. 재조합 바이러스의 배양세포주에서 외래 유전자 발현에 대한 누에 체액의 첨가 효과는 FBS가 2% 또는 5% 일때 누에 체액 2%의 소량 첨가만으로도 누에체액이 첨가되지 않았을 때에 비해 약 2배 이상 높은 발현량을 확인하였으며, 그 보다 FBS가 전혀 첨가되지 않고 단지 누에 체액 2% 첨가만으로도 FBS 첨가시와 마찬가지의 높은 발현량을 보여 누에 체액에 의한 FBS의 대체 효과를 나타내었다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권6호
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• pp.989-993
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• 2007

A lactose-hydrolyzed milk with low sweetness was developed using nanofiltration. Raw milk was treated with 0.03% ${\beta}$-galactosidase at $4^{\circ}C$ for 24 h to hydrolyze lactose partially. The resultant lactose-hydrolyzed milk containing 0.43% lactose was then concentrated using a nanofiltration membrane to reach concentration factor of 2.13. The concentration factors and coefficients of retention of milk components in nanofiltration were determined. The concentration factor of milk fat was 2.20 which was the highest of the milk components. The coefficient of retention of calcium and riboflavin was 0.96 and 0.76, respectively. However, the coefficient of retention of glucose, galactose, and sodium was 0.21, 0.15, and 0.22, respectively. Raw milk was treated with 0.1% ${\beta}$-galactosidase at $4^{\circ}C$ for 40 h to hydrolyze lactose fully and then concentrated to reach a concentration factor of 1.6 by using nanofiltration. The concentrated milk was reconstituted with water. The lactose-hydrolyzed milk had sweetness similar to milk. The compositional ratios of crude protein, calcium, sodium, and riboflavin of lactose-hydrolyzed nanofiltrated milk to those of raw milk were 99%, 97%, 77%, and 80%, respectively. This study showed that nanofiltration of lactose-hydrolyzed milk to remove galactose and glucose did not cause significant loss of calcium. The lactose-hydrolyzed nanofiltrated milk contained 0.06% lactose and had sweetness similar to milk.