• Asian-Australasian Journal of Animal Sciences
• /
• 제13권12호
• /
• pp.1770-1774
• /
• 2000

Extended use of porcine blood in food ingredients depends on the decolorization of red blood cell concentrates and the modification of its functional properties. The purpose of this study is to compare the relative effect of cation ion exchanger for decolorization of porcine red blood globin. The globin extract is freeze-dried for determination of various functional properties, such as solubility, emulsion capability and foaming ability. Since the isoelectric point of blood globin is located at pH 6.8, which is the neutral pH ranges (6-8), so its functionalities are inferior around these pHs. This weakness has been the main reason, which limit the extended use of blood globin in food industry. Acetylation and succinylation of blood globin can be an alternative way to improve its functionalities. These results may provide new information to understand the decolorization mode by cation ion exchanger for the blood globin. With chemical, the functionalities of blood globin could be obviously improved. The above findings could enable food industry to extend the use of blood globin as a food ingredient.

• 생명과학회지
• /
• 제8권3호
• /
• pp.249-256
• /
• 1998

일본의 잡종잉어 ${\beta}$사 globin유전자의 염기배열로부터 추정되는 아미노산 배열과 이미 보고되어 있는 ${\beta}$사 globin의 아미노산 배열을 비교한 결과, 금붕어와 가장 높은 상동성인 97.3%를 나타내었으며, 거울잉어 ${\beta}_B$와 95.2%, 거울 잉어 ${\beta}_A$와 93.9%의 높은 상동성을 나타내었다. 다음으로 송어(76.9%), 전기뱀장어(71.4%), 혹다랑어(61.9%), 성대(59.9%), 양태(58.5%), 시라칸스(52.4%), 폐어(46.9%), 상어(38.1%) 및 전기가오리(36.1%)의 순으로 나타났다. 또한 ${\beta}$사 globin의 아미노산 배열의 상동성을 기초로하여 분자진화의 계통수를 구하였을 때, 각각의 진화거리로부터 일본의 잡종잉어와 금붕어가 0.013으로 가장 가깝고, 분기한 것이 가장 근년인 것으로 생각되어졌다. 그 다음이 같은종인 거울잉어로 진화거리는 0.035였으며, 연골어류나 폐어와는 그 진화거리가 크게 벌어져 있어 상당히 오래전에 분기한 것으로 추정되었다. 일본 잡종이어의 ${\beta}$사 globin유전자로부터 추정되는 아미노산 배열과 이미 보고되어 있는 다른 척추동물의 아미노산 배열과 비교한 결과, 상동성은 오리(58.1%), 닭(57.8%), 쥐(55.1%), 사람(52.4%), 개구리(50.7%), 및 염소(45.9%)의 순으로 나타났다.

• 대한의생명과학회지
• /
• 제11권1호
• /
• pp.85-88
• /
• 2005

Hemoglobin is oxygen carrier protein within erythrocyte in blood. Apoprotein of this, globin, is synthesized in the cytosol but it's cofactor, heme, is synthesized in the mitochondria. It has not been known very well how globin receives the heme from mitochondria and folds to hemoglobin. In this folding process, the initial structure of globin seems to be very important. A small volume of globin at acid pH was added rapidly into the bulk of an egg phosphatidylcholine $60\%$ liposome, containing hemins, at neutral pH according to the Rapid Dilution method. It was observed that an acid-induced unfolding structure of globin is initially needed to receive hemins from the lipid bilayer of liposomes. Also, this conclusion was confirmed with the absorption spectrum of the refolded globin separated by centrifugation.

• 대한의생명과학회지
• /
• 제11권3호
• /
• pp.349-353
• /
• 2005

The globin in cytosol receives the heme in mitochondria and folds to the hemoglobin within erythrocyte. Two mechanisms have been proposed that the heme was transfered post-translationally or cotranslationally to the globin. In this research, how the globin in cytosol receives post-translationally the heme from membranes was studied according to pH and phospholipid composition. Globins dissolved in various pH buffer solutions $(PH\;3\~7)$ were rapidly added into the bulk of egg phosphatidylcholine $100\%\;or\;60\%$ liposomes containing hemin in pH 7 buffer solution. Hemin was very highly transferred to globin at pH 4 and 6. Also, hemin was more efficiently transfered to globin in egg phosphatidylcholine $100\%$ than in $60\%$ liposomes.

• 생명과학회지
• /
• 제8권3호
• /
• pp.348-351
• /
• 1998

Common DNA fragments of the ${\beta}$-globin gene were observed from six races of the adult common carp: Hybrid-Yamato, Japanese wild type, Mirror, Suwa-Yamato, Scale German, and Saku-Yamato. Chromosomal DNAs isolated from the above six races were digested with restriction endonucleased EcoRI and PstI. The digested fragments were transferred onto nitrocellulose filter and hybridized with a probe of carp ${\beta}$-globin cDNA. Molecular sizes of the hybridized DNA fragments digested with EcoRI were 3.6Kb(Kilo base), 4.3Kb and 15Kb in Hybrid-Yamato, Japanese wild type, Mirror, Scale German and Saku-Yamato carp DNAs. In Scale German and Saku-Yamato carp DNAs, two and one more hybridized DNA fragments were observed, respectively. Molecular sizes of the hybridized DNA fragments digested with PstI were 2.2Kb, 6.5Kb, 7.8Kb and 9.2Kb in Hybrid-Yamato, 2.2Kb, 6.5Kb and 9.2Kb in Japanese wild type, 2.2Kb, 6.5Kb, 7.8Kb, and 13Kb in Mirror, 2,2Kb, 5,5Kb, 6.5Kb, 7.8Kb, 9.2Kb and13Kb in Scale German, and 2.2Kb, 5.5Kb, 6.5Kb, 9.2Kb and Saku-Yamato carp DNA. Therefore, depending on carps, three to six DNA fragments were hybridized with ${\beta}$-globin gene probe. Thus it indicated polymorphysm in the globin gene family of carp.

• Bulletin of the Korean Chemical Society
• /
• 제23권8호
• /
• pp.1073-1077
• /
• 2002

Binding of dodecyl trimethylammonium bromide (DTAB) to human and bovine hemoglobin and globin samples has been investigated in 50 mM glycine buffer pH = 10, I = 0.0318 and 300 K by equilibrium dialysis and temperature scanning spectrophotometry techniques and method for calculation of average hydrophobicity. The binding data has been analyzed, in terms of binding capacity concept $({\theta})$, Hill coefficient (nH) and intrinsic Gibbs free energy of binding $({\Delta}Gbv).$ The results of binding data, melting point (Tm) and average hydrophobicity show that human hemoglobin has more structural stability than bovine hemoglobin sample. Moreover the results of binding data analysis represent the systems with two and one sets of binding sites for hemoglobin and globin, respectively. It seems that the destabilization of hemoglobin structure due to removal of heme group, is responsible of such behavior. The results indicating the removal of heme group from hemoglobin caused the depletion of first binding set as an electrostatic site upon interaction with DTAB and exposing the hydrophobic patches for protein.

• 한국동물학회지
• /
• 제27권2호
• /
• pp.103-116
• /
• 1984

유전자 구조 및 유전정보 흐름의 차이로 인하여 고등생물의 유전자를 미생물에 직접 cloning하면 원하는 유전자 산물을 얻지 못하는 경우가 많다. 이것을 극복하기 위해서는 화학적인 방법으로 유전자를 합성하든지, 또는 역제효소를 사용하여 고등생물의 mRNA로부터 유전자를 합성하여 cloning하는 방법을 사용한다. 본 연구에서는 oligo(dT)-cellulose column 방법으로 순수분리한 plasmid pBR322의 Pst I site에 cloning하였다. 우선 AMV reverse transcriptase로 primary cDNA를 합성하고, 알칼리를 처리하여 주형 RNA를 제거했다. 이번에는 이 primary cDNA를 주형으로 Klenow enzyme과 reverse transcriptase를 차례로 처리하여 double stranded DNA를 합성하고, 이 때 5' end 근처에 형성되는 hairpin loop을 Sl nuclease로 제거했다. Terminal deoxynucleotidyl transferase를 사용하여, 합성된 dsDNA에는 poly(dC) track을, Pst I endonuclease를 처리한 plasmid DNA에서는 poly(dG) track을 각각 붙인다음 이들을 서로 annealing시키고 E. coli에 transformation시켜서 크기가 큰 plasmid를 갖는 clone을 cracking 방법으로 일처 선별하였다. 이렇게 선별된 clone을 in 냐셔 hybridization 방법으로 조사하여 globin DNA가 들어간 colony를 이차 선별하고 여러 restriction enzyme으로 잘라보아 globin DNA가 cloning된 것을 확인하였다. 토끼 hemoglobin으로 immunize한 rat (Wistar)에서 뽑은 제일차 혈청과 염소에서 뽑은 제이차 혈청의 antibody를 사용한 radioimmunoassay방법으로, cloning된 globin gene이 대장균내에서 발현되는 지의 여부를 살펴 보았는데, 박테리아의 $\\beta$-lactamase와 토끼의 globin이 결합된 chimeric protein이 대장균 내에서 다량 합성되며, 이 단백질은 토끼 hemoglobin의 antigenic determinant를 가지고 있음을 알 수 있었다.

• BMB Reports
• /
• 제42권8호
• /
• pp.493-499
• /
• 2009

MicroRNAs (miRs) are a family of small noncoding RNAs that regulate gene expression by targeting mRNAs in a sequence specific manner, inducing translational repression or mRNA degradation. In this paper we have first analyzed by microarray the miR-profile in erythroid precursor cells from one normal and two thalassemic patients expressing different levels of fetal hemoglobin (one of them displaying HPFH phenotype). The microarray data were confirmed by RT-PCR analysis, and allowed us to identify miR-210 as an highly expressed miR in the erythroid precursor cells from the HPFH patient. When RT-PCR was performed on mithramycin-induced K562 cells and erythroid precursor cells, miR-210 was found to be induced in time-dependent and dose-dependent fashion, together with increased expression of the fetal $\gamma$-globin genes. Altogether, the data suggest that miR-210 might be involved in increased expression of $\gamma$-globin genes in differentiating erythroid cells.

• 한국환경보건학회지
• /
• 제30권4호
• /
• pp.301-307
• /
• 2004

1,3-butadiene(DB) is a well-established rodent carcinogen, and a probable carcinogen to humans. This study was investigated the formation of hemoglobin adduct in ICR female mice inhaled with BD for 3 weeks (5 hr/day, 5 days/week). Body weights of mice were significantly low from onward day 4 or 9 of exposure comparing the control. Hemoglobin adducts formed by DB exposure were (N-2-hydroxy-3-butenyl) valine (HB Val adduct) and (N-2,3,4-trihydroxy-butyl)valine(THB Val adduct). The levels of HB Val adducts were 1.8, 3.7 and 6.2 pmol/mg globin for the 500 ppm BD inhalation group, and 5.7, 7.4 and 16.0 pmol/mg globin for the 1000 ppm BD inhalation group, when observed on the $1^{st},\;2^{nd},\;and\;3^{rd}$ week after inhalation exposure, respectively. The levels of THBVal adducts were 32.0, 42.0 and 55.0 pmol/mg globin for the 500 ppm DB inhalation group, and 67.8, 72.7 and 83.5 pmol/mg globin for the 1000 ppm BD inhalation group, when observed on the $1^{st},\;2^{nd},\;and\;3^{rd}$ week after inhalation exposure, respectively. Ratios of THBVal and HBVal adducts were higher at earlier exposure period and lower concentration. Ratios on the $1^{st},\;2^{nd},\;and\;3^{rd}$ week were 17.8, 11.4 and 8.87 for the 500 ppm BD inhalation group, and 11.9, 9.8 and 5.2 for the 1000 ppm BD inhalation group, respectively. In conclusion, THB Val and HB Val adducts were the important hemoglobin adducts in ICR female mice inhaled with BD, and the latter was more appropriate biomarker than the other for biological monitoring of BD exposure.

• 한국환경보건학회:학술대회논문집
• /
• 한국환경보건학회 2004년도 International Conference Global Environmental Problems and their Health Consequences
• /
• pp.73-86
• /
• 2004

The purpose of this study is the identification of (N-2-hydroxy-3-butenyl) valine(HBVal adduct) and (N-2,3,4-trihydroxy-butyl)valine(THBVal adduct)with mice inhalation exposure with 1,3-butadiene for 3 weeks($6\;hr/day\;{\times}\;5\;days/week$). Body weights were significantly lower from 4 or 9 exposure post-day in 1000 or 500ppm inhalation group than in control. The levels of HBVal adducts are 1.8, 3.7 and 6.2 pmol/mg globin in $1^{st}$, $2^{nd}$ , and $3^{rd}$ week for 500 ppm 1,3-butadiene(BD), and 5.7, 7.4 and 16.0 pmol/mg globin in $1^{st}$, $2^{nd}$ , and $3^{rd}$ week for 1000 ppm BD inhalation exposure. The levels of THBVal adducts are 32.0, 42.0 and 55.0 pmol/mg globin in $1^{st}$, $2^{nd}$ , and $3^{rd}$ week for 500 ppm BD, and 67.8, 72.7 and 83.5 pmol/mg globin in $1^{st}$, $2^{nd}$ , and $3^{rd}$ week for 1000 ppm BD inhalation exposure. Their ratios of THBVal and HBVal adducts are higher at earlier exposure and lower concentration. They are17.8, 11.4 and 8.87 in $1^{st}$, $2^{nd}$ , and $3^{rd}$ week for 500 ppm BD, and 11.9, 9.8 and 5.2 in $1^{st}$, $2^{nd}$ , and $3^{rd}$ week for 1000 ppm BD inhalation exposure. In conclusion, THBVal and HBVal adducts are a important hemoglobin adduct for monitoring of BD exposure, and the latter is more biomarker than the other.