Neurotensin was found present in human early placenta using RIA and its release was demonstrated by in vitro perfusion experiment.Immunohistochemical staining revealed lots of NT-positive granules in early placental ...Neurotensin was found present in human early placenta using RIA and its release was demonstrated by in vitro perfusion experiment.Immunohistochemical staining revealed lots of NT-positive granules in early placental villi. Functional testing displayed NT′s inhibitory effect on release of hCG and progesterone from incubated early placenta. This result suggests the potential use of NT as an agent for terminating early pregnancy.展开更多
Recently,some of the genetic mechanisms of heart specification have been elucidated in Drosophila .However,genes involved in early cardiogenesis of human remain to be identified.Since the pathways that regulate ear...Recently,some of the genetic mechanisms of heart specification have been elucidated in Drosophila .However,genes involved in early cardiogenesis of human remain to be identified.Since the pathways that regulate early cardiac fate determination are conserved between Drosophila and vertebrates,flies can be used as a model test system to explore the genetic basis of cardiogenesis in human.In this project,about 3000 reccieve lethal gene lines were produced by P or EMS mutagenesis.With staining of antibodies against heart precussor cells of Drosophila ,about 200 lines were observed to show heart phenotype.In pilot studies of their function with RNAi technique,the RNAi phenotypes of several genes tested were observed,which were very similar to that of their mutants,showing heart tube defects or no heart precursors formation.Taking advantage of the advanced genetic information available in the Drosophila and human systems,we have identified about 50 human transcripts homologous to the Drosophila heart related gene candidates.Northern blot analysis for some of the human candidates showed that several genes were expressed in both adult and early embryonic tissues,which may help in the evaluation of candidate genes for human cardiogenesis.Our further experiments with transgenic flies generated with wild type and mutant forms of these candidate genes to examine for defects in cardiogenesis or cardiac function are under way.The candidate genes producing cardiac specific defects suggestive of similarities to the heart disease syndromes can then be pursued further as likely disease gene candidates.Such an approach is likely to provide a dramatic reduction of possible candidate genes,or to screen and identify mutations that may generate the disease in human.展开更多
The Primacy question addressed in our study is: Is the difterntial expression of rbcL gene in mesophyll cells and in bundle sheath cells related to the sequence of the gene per se?An enzymatic approach was fist establ...The Primacy question addressed in our study is: Is the difterntial expression of rbcL gene in mesophyll cells and in bundle sheath cells related to the sequence of the gene per se?An enzymatic approach was fist established to separate the two groups of cells. Microscopic examination revealed satisfactory separation effect: minimal mutual contamination was found so that no mistake might be introduced into biochemical or molecular biological expeitments using such preparations. CpDNA were isolated from mesophyll cells and from bundle sheath cells and coding region of rbcL gene was obtained from each by PCR ampilfication.Cloning and sequencing were then done on them.Compartive analysis , however, revealed identical sequence, with a length of 1,368 bp, encoding 456 amino acids. Since sequences of the non-coding regions of rbcL gene in masephyll sad bundle sheath have not been obtained, it can not yet be concluded that the differential expression is not related to the sequence itself. Nevertheless,It sesems justifiable to infer that whatever difference there may be between the sequences of rbcL gene in two groups of cells can only be found in the non-coding regions(including promoter and the 3' down stream region).展开更多
Mus musculus tyrosine 3-monooxygenase/trytophan 5-monooxygenase activation protein,zeta polypeptide(Ywhaz)is related to communication among blastomeres and con-tributes to buildup of celiular communication system,and ...Mus musculus tyrosine 3-monooxygenase/trytophan 5-monooxygenase activation protein,zeta polypeptide(Ywhaz)is related to communication among blastomeres and con-tributes to buildup of celiular communication system,and ATPase 6 is essential for one-cell to two-cell transition and play an important role in establishment of oxidative phosphorylation.We report the expression analysis of mouse two-celi stage specific genes using modified mR-NA differential display.The results of sequence analysisand reverse northern blot indicate that the genes for ATPase 6and Ywhaz are expressed just in 2-cell embryo.展开更多
A new method is presented with which we isolated milochondrial DNA from fresh carp liver usingdifferential centrifugation and DNase treatment that gave high yield of purified product with an easyand economical procedu...A new method is presented with which we isolated milochondrial DNA from fresh carp liver usingdifferential centrifugation and DNase treatment that gave high yield of purified product with an easyand economical procedure. Highly distinct bands were displayed in agarose gel electrophoresls ofthe product digested with restrictlon enzymes, which were successfully used in constructingrestriction map and molecular clone of mitochondrial genes. With DNAs thus obtained, we havecloned cysteine tRNA gene (tRNA^(Cys) gene) of carp mitochondria, determined the nucleotide sequenceof it and the light strand origin, and depicted the cloverleaf secondary structure of tDNA^(Cya) and thelight strand origin. Analysis of nucleotide sequences of tRNA^(Cy) genes of 5 vertebrates has revealedunusual features of carp mitochondrial tRNA^(Cy) gene as compared with their cytoplasmic counter-parts, Altogether 36 bases were found in the light strand origin of carp mitochondriaf: 11 pairs in thestem; and 14 bases in the loop. As compared with those of other 11 vertebrate species, the sequenceof the stem is very conservative while both sequence and length of the loop are quite variable. Thestructure of the stem-loop may play an important role in light strand replication.展开更多
Sperms from Pink salmon were subjected to incomplete irradiation by γ-ray to cause partial breakageof their chromosome. Normal eggs from masu salmon were fertilized by these damaged sperms andput under hydrostatic pr...Sperms from Pink salmon were subjected to incomplete irradiation by γ-ray to cause partial breakageof their chromosome. Normal eggs from masu salmon were fertilized by these damaged sperms andput under hydrostatic pressure to suppress the release of second polar bodies. The resultant eggscontained complete sets of genome from masu salmon and some chromosomes and chromosomefragments from pink salmon. Karyotype and isozyme of the embryos were analyzed. The resultsdemonstrate that the chromosomes from pink salmon showed genetic activities. Variations in hatchedindividuals were observed.展开更多
文摘Neurotensin was found present in human early placenta using RIA and its release was demonstrated by in vitro perfusion experiment.Immunohistochemical staining revealed lots of NT-positive granules in early placental villi. Functional testing displayed NT′s inhibitory effect on release of hCG and progesterone from incubated early placenta. This result suggests the potential use of NT as an agent for terminating early pregnancy.
文摘Recently,some of the genetic mechanisms of heart specification have been elucidated in Drosophila .However,genes involved in early cardiogenesis of human remain to be identified.Since the pathways that regulate early cardiac fate determination are conserved between Drosophila and vertebrates,flies can be used as a model test system to explore the genetic basis of cardiogenesis in human.In this project,about 3000 reccieve lethal gene lines were produced by P or EMS mutagenesis.With staining of antibodies against heart precussor cells of Drosophila ,about 200 lines were observed to show heart phenotype.In pilot studies of their function with RNAi technique,the RNAi phenotypes of several genes tested were observed,which were very similar to that of their mutants,showing heart tube defects or no heart precursors formation.Taking advantage of the advanced genetic information available in the Drosophila and human systems,we have identified about 50 human transcripts homologous to the Drosophila heart related gene candidates.Northern blot analysis for some of the human candidates showed that several genes were expressed in both adult and early embryonic tissues,which may help in the evaluation of candidate genes for human cardiogenesis.Our further experiments with transgenic flies generated with wild type and mutant forms of these candidate genes to examine for defects in cardiogenesis or cardiac function are under way.The candidate genes producing cardiac specific defects suggestive of similarities to the heart disease syndromes can then be pursued further as likely disease gene candidates.Such an approach is likely to provide a dramatic reduction of possible candidate genes,or to screen and identify mutations that may generate the disease in human.
文摘The Primacy question addressed in our study is: Is the difterntial expression of rbcL gene in mesophyll cells and in bundle sheath cells related to the sequence of the gene per se?An enzymatic approach was fist established to separate the two groups of cells. Microscopic examination revealed satisfactory separation effect: minimal mutual contamination was found so that no mistake might be introduced into biochemical or molecular biological expeitments using such preparations. CpDNA were isolated from mesophyll cells and from bundle sheath cells and coding region of rbcL gene was obtained from each by PCR ampilfication.Cloning and sequencing were then done on them.Compartive analysis , however, revealed identical sequence, with a length of 1,368 bp, encoding 456 amino acids. Since sequences of the non-coding regions of rbcL gene in masephyll sad bundle sheath have not been obtained, it can not yet be concluded that the differential expression is not related to the sequence itself. Nevertheless,It sesems justifiable to infer that whatever difference there may be between the sequences of rbcL gene in two groups of cells can only be found in the non-coding regions(including promoter and the 3' down stream region).
基金The project was supported by the State Basic Research Development Program of“973”(No.G200016107)
文摘Mus musculus tyrosine 3-monooxygenase/trytophan 5-monooxygenase activation protein,zeta polypeptide(Ywhaz)is related to communication among blastomeres and con-tributes to buildup of celiular communication system,and ATPase 6 is essential for one-cell to two-cell transition and play an important role in establishment of oxidative phosphorylation.We report the expression analysis of mouse two-celi stage specific genes using modified mR-NA differential display.The results of sequence analysisand reverse northern blot indicate that the genes for ATPase 6and Ywhaz are expressed just in 2-cell embryo.
基金This work was supported by grant from National Foundation of Natural Sciences of China.
文摘A new method is presented with which we isolated milochondrial DNA from fresh carp liver usingdifferential centrifugation and DNase treatment that gave high yield of purified product with an easyand economical procedure. Highly distinct bands were displayed in agarose gel electrophoresls ofthe product digested with restrictlon enzymes, which were successfully used in constructingrestriction map and molecular clone of mitochondrial genes. With DNAs thus obtained, we havecloned cysteine tRNA gene (tRNA^(Cys) gene) of carp mitochondria, determined the nucleotide sequenceof it and the light strand origin, and depicted the cloverleaf secondary structure of tDNA^(Cya) and thelight strand origin. Analysis of nucleotide sequences of tRNA^(Cy) genes of 5 vertebrates has revealedunusual features of carp mitochondrial tRNA^(Cy) gene as compared with their cytoplasmic counter-parts, Altogether 36 bases were found in the light strand origin of carp mitochondriaf: 11 pairs in thestem; and 14 bases in the loop. As compared with those of other 11 vertebrate species, the sequenceof the stem is very conservative while both sequence and length of the loop are quite variable. Thestructure of the stem-loop may play an important role in light strand replication.
文摘Sperms from Pink salmon were subjected to incomplete irradiation by γ-ray to cause partial breakageof their chromosome. Normal eggs from masu salmon were fertilized by these damaged sperms andput under hydrostatic pressure to suppress the release of second polar bodies. The resultant eggscontained complete sets of genome from masu salmon and some chromosomes and chromosomefragments from pink salmon. Karyotype and isozyme of the embryos were analyzed. The resultsdemonstrate that the chromosomes from pink salmon showed genetic activities. Variations in hatchedindividuals were observed.
