• The Korean Journal of Zoology
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• v.35 no.1
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• pp.8-16
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• 1992

포유류의 심방에 주로 존재하여 체액 및 전해질 대사의 조절과 혈압 조절에 관여하는 atrial natriuretic peptide(ANP)가 담수어류인 미꾸라지(Misgumus mizolepis G.)의 심장 및 기타조직에도 존재하는지 알아보고 이들의 분자적 특성 및 온도차에 의한 함량변동을 조사하였다. 미꾸라지의 심장과 동맥구의 추출액은 모두 희석농도에 따라 표준곡선에 평행하는 ANP immunoreactivity를 나타냈다. 미꾸라지의 심장과 동맥구 모두에서 저분자량 및 고분자량의 immunoreactive ANP(irANP)가 관찰되었다. 한편 미꾸라지의 심장내 irANP함량은 61.56 $\pm$ 5.08 pg/mg이었으며, 심장외 조직인 동맥구는 60.59 $\pm$ 5.04 pg/mg으로 거의 동량의 irANP를 함유하였다. 미꾸라지를 저온상태에 적응시킨 경우에는 심장보다도 동맥구에서 더 유의한 irANP의 함량변동을 보였다(p < 0.01), 미꾸라지의 심장 및 동맥구에 존재하는 irANP는 주로 고분자량의 Pro-ANP가 관찰되므로써 미꾸라지의 심장과 동맥구 모두가 irANP를 조직내에서 합성할 것임을 시사하였다. 아울러 온도는 미꾸라지의 경우에서 irANP의 함량변동에 영향을 주는 요인임을 알 수 있었다.

• The Korean Journal of Zoology
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• v.37 no.3
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• pp.452-465
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• 1994

미꾸리속의 미꾸리(Misgumus anguillicaudatus)와 미꾸라지(M. mizolepis)의 종내 및 종간 유전적 변이와 유전적 차이를 알아보고자 2종 15개집단과 대만산 Paramisgumus김늠bwanus 1개 집단을 대상으로 전기영동법의 의한 동위효소분석을 실시한 결과 각. ongui각caudotus 9개 집단의 평균 유전적 변이정도는 P=34.20%, Ho=0.099, He=0 114였고. M. mizolepgs 6개 집단은 평균 P=35.55%, Ho=0 141, He=0.148이었으며, p연abwonus는 P=45.9%, Ho=0.132. 연e=0.119로서 일반적인 담수어류에 비해 높은 변이를 나타내었다 종간 평균 유전적 근연치를 비교한 결과 M. aneuflISca udatuo와 M. mfzole부랍 사이는 5=0.467이었고. M. angu연흘audatus와 P. dabrvanus 사이는 5=0.475로 비교적 근연관계가 낮았으나 M. mizolep결와 P. dabwanuo 사이는 5=0.834로 매우 가까운 유연관계를 나타냈다 유전적 차이치를 토대로 M. ongu비상라udctus와 M. mfzolep결의 종분화 연대를 산출한 결과 이들은 약 350만년전에 분화된 것으로 추정되었다.

• Journal of Aquaculture
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• v.8 no.3
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• pp.209-220
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• 1995

The nuclear matrix was isolated from Misgumus mizolepis liver nuclei by low salt extraction and restriction enzyme treatment. The structure was digested with proteinase K. After centrifugation, matrix attachment regions (MARs) were obtained by RNase treatment and phenol-chloroform extraction. The result leads to the appearance of smeared bands in the range of about 0.3-15 kb. pURY19 vector was constructed by inserting 2.13 kb Eco47 III fragment of the yeast uracil 3 gene into the unique Ssp I site of pUC19 plasmid vector as a selection marker. This vector is unable to be maintained in Sacrharomyces cerevisiae by itself since it cannot replicate as an extrachromosomal element. Using this system, we attempted cloning the ARS (autonomously replicating sequence) from M. mizelepis to develop an efficient expression vector for the transgenic fish. pURY19N_{l-62}$ were constructed by inserting MARs in pURY19 plasmid vector and transformation of E. coli $DH5\alpha$. Replication origins (ARS) of M. mizolepis were isolated, which enabled the vector to replicate autonomously in S. cerevisiae. The cloned DNA fragments were sequenced by Sanger's dideoxy-chain termination method. All clones were AT-rich. $pURY19N_6$, one of the clones, expecially contained ARS consensus sequence, Topoisomerase II consensus, near A-box and T-box.

• Journal of fish pathology
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• v.18 no.1
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• pp.67-80
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• 2005

Phosphoinositide-specific phospholipase $C\delta$ $PLC\delta$) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to obtain the biochemical characteristics of the expressed recombinant $PLC\delta$ in E. coli cloned from Misgurnus mizolepis and partially purified $PLC\delta$ enzymes from liver tissues of M. mizolepis (wild ML-$PLC\delta$). The ML $PLC\delta$ gene was cloned and expressed under the previous report (Kim et al., 2004), and purified the recombinant protein by successive chromatography using $Ni^{2+}$-NTA affinity column and gel iltration FPLC column. The wild ML-$PLC\delta$ protein was solublized with 2 M KCI and purified by successive chromatography on open heparin-Sephagel and analytical TSKgel heparin-5PW. Both the recombinant and wild ML-$PLC\delta$ form of protein showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bis-phosphate (PIP$_2$) or phosphatidylinositol (PI). Its activity was absolutely $Ca^{2+}$- dependant, which was similar to mammalian $PLC\delta$ isozymes. Maximal PI-hydrolytic activations of recombinant and wild ML- TEX>$PLC\delta$ was at pH 7.0 and pH 7.5, respectively. In addition, the enzymatic activities of recombinant and wild ML-$PLC\delta$ were increased in concentration-dependent manner by detergent, such as sodium deoxycholate SDC), phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The activities decreased in contrast by a polyamine, such as spermine. Western blotting showed that several types of $PLC\delta$ isozymes exist in various organs. Taken together our results, it suggested that the biochemical characteristics of ML-$PLC\delta$ are similar with those of mammalian $PLC\delta1$ and ${\delta}3$ isozymes.