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Yan-Hong He Guo-Gui Ning Ya-Lin Sun Yan Hu Xing-Yu Zhao Man-Zhu Bao 《Molecular breeding : new strategies in plant improvement》2010,26(1):19-29
A male sterile line was isolated in marigold (Tagetes erecta L.) and cytological analysis determined this to be a novel genic male sterility trait (Tems). Through the use of amplified fragment length polymorphisms (AFLPs) and bulked segregant analysis (BSA), tightly linked
markers of Tems were identified with a view towards a map-based cloning strategy. It was found that spontaneous homeotic conversion of floral
organs was the underlying cause of the male sterility in this marigold line. Thus, petals of male sterile plants resembled
sepal-like structures and the stamens were partially converted to styles, although without the full characteristics or function
of the true style organs. We have constructed a fine marker-based map for the Tems gene. This is intended to provide a tool for marker assisted selection (MAS) strategies in hybrid breeding and map-based
cloning strategies for the male sterility locus. We discuss the significance of this spontaneously derived genic male sterility
trait relating to the homeotic conversion of floral organs in marigold. 相似文献
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15.
Xue-Yuan Dong Yun-Yan Li Xiao-Ang Yang Wei-Feng Chen 《Biochimie et biologie cellulaire》2004,82(5):577-582
In an effort to identify novel Cancer-Testis genes, we analyzed the sequence in the q26-28 region of human X chromosome by several on-line tools. The candidate sequences were then confirmed by experiments. We have obtained a novel Cancer-Testis gene, BJ-HCC-20. In vivo, it was found to have two isoforms. In samples of liver, colon, gastric and lung cancer tested, the expression frequency of BJ-HCC-20 is 25%, 17%, 21% and 15%, respectively. Full-length cDNAs of both BJ-HCC-20 isoforms were isolated and their gene structures and promoter regions were characterized. BJ-HCC-20 might have implications in theoretical and practical tumor biology. 相似文献
16.
An endopeptidase (LEP-II), which has a unique substrate specificity, was purified to homogeneity by conventional chromatographic techniques from Streptococcus cremoris H61. The enzyme was a metalloendopeptidase since it was inhibited by EDTA and 1,10-phenanthroline; the metal-depleted enzyme could be fully reactivated by micromolar levels of Zn2+ and was not inhibited by specific inhibitors for serine or thiol protease. The molecular mass of the enzyme was estimated to be 80 kDa by Sephacryl S-300 gel filtration and high-performance liquid chromatography with a TSK-G3000SW column. The enzyme consisted of two identical subunits and the N-terminal sequence of LEP-II was determined up to the 19th residue. Although the enzyme had a broad substrate specificity it specifically hydrolyzed the peptide bonds involving the amino groups of hydrophobic amino acid residues. Various small polypeptides, such as alpha s1-CN(f1-23), alpha s1-CN(f91-100), oxidized insulin B chain, glucagon and some biologically active peptides were hydrolyzed. However, a variety of larger polypeptides or proteins, such as alpha s1-CN(f1-54), alpha s1-CN(f61-123), alpha s1-CN(f136-196), alpha s1-casein, beta-casein, and kappa-casein were not hydrolyzed. LEP-II recognized the size of its substrates, which were limited below a molecular mass of about 3.5 kDa. 相似文献
17.
Expression of an enhancer-binding protein in insect cells transfected with the Autographa californica nuclear polyhedrosis virus IE1 gene. 总被引:10,自引:10,他引:0
The baculovirus Autographa californica nuclear polyhedrosis virus contains an element known as homologous region 5 (hr5) which is an enhancer of delayed-early viral gene expression. To begin to identify proteins that interact with hr5, DNA-protein interactions were analyzed by using extracts from Spodoptera frugiperda cells and a fragment of DNA containing the left half of the hr5 enhancer. This 252-bp DNA fragment contains two copies of a 30-bp direct repeat (DR30) and two copies of a 24-bp imperfect palindrome contained within a 60-bp direct repeat (DR60). Extracts prepared from normal S. frugiperda cells and cells transfected with pUC8 lacked enhancer-binding proteins. However, when gel shift assays were performed with extracts from cells transfected with a plasmid containing the viral trans-activator IE1 gene, two DNA-protein complexes were formed. Both DNA-protein complexes were specifically inhibited by competition with a 60-bp oligonucleotide corresponding to DR60 but not by competition with a different oligonucleotide corresponding to DR30. Formation of the two complexes did not appear to involve cooperative interactions between binding proteins. When DR60 was used as a probe, a single complex was formed. To measure the enhancer activity of DR60, a reporter plasmid was constructed that contained DR60 cloned upstream of the reporter chloramphenicol acetyltransferase gene under the control of the delayed-early 39K promoter. Transient expression analysis indicated that the oligonucleotide increased expression of this gene 300-fold over the level obtained in the absence of any enhancer sequences. 相似文献
18.
P Folk J Dong J L Wittlife 《The Journal of steroid biochemistry and molecular biology》1992,42(2):141-150
Polymorphism of estrogen (ER) and progestin receptors (PR) was analyzed simultaneously using high performance hydrophobic interaction chromatography (HPHIC). HPHIC was used previously to characterize four ER isoforms [Hyder et al., J. Chromat. 397 (1987) 251] based on retention times on Synchropak propyl (100 x 6 mm) HPLC columns (Synchrom, Inc.). ER and PR were prepared from human breast cancer. ER was labeled with 3 nM of either [3H]estradiol-17 beta ([3H]E) or [125I]iodoestradiol-17 beta ([125I]E) while PR was associated with 5 nM of either [3H]R5020 ([3H]R) or [125I]iodovinylnortestosterone ([125I]V). ER was resolved by HPHIC into isoforms MI (Rt = 11 min), I(Rt = 16 min), and II (Rt = 24 min). Isoforms I and II each accounted for ca 45% of specific binding. PR separated into isoforms MI (Rt = 14 min) and I (Rt = 21 min, 80% of specific binding) when eluted with the same gradient used for ER chromatography. Upon inclusion of 10 mM molybdate ER resolved into isoforms MI and MII (Rt = 16 min) and PR into isoforms MI and I (here however isoform MI represented 80-95% of specific binding). Elution patterns were preserved with different batches of stationary phase suggesting the integrity of the isoform distribution. HPLC profiles of ER isoforms labeled with earlier [125I]E or [3H]E were identical as were PR isoform profiles labeled with either [3H]R or [125I]V. Pairs of 125I- and 3H-labeled ligands were used in either combination to monitor ER and PR profiles simultaneously. Isoforms analyzed in 50 biopsies gave reproducible retention times, however the ratio between I and II for ER and MI and I for PR varied. This method allows rapid, simultaneous monitoring of the chromatographic behavior of ER and PR isoforms or other associating proteins or nucleotides. One may now better elucidate their interrelationship as it relates to the hormone-response mechanism. 相似文献
19.
S Y Yan Z R Mao H Y Yang M A Tu S H Li G P Huang G S Li L Guo G Q Jin R F He 《The International journal of developmental biology》1991,35(4):429-435
The nucleus of a blastula cell from Tilapia (Oreochromis nilotica, family Cichlidae, order Perciformes) was transplanted into an enucleated egg of Loach (Paramisgurnus dabryanus, family Cobitidae, order Cypriniformes). From among 3747 nucleo-cytoplasmic hybrid (NCH) eggs two NCH larval fish (0.05%) were obtained; one died on the 6th day and the other died on the 12th day after the operation. Morphological examinations showed that both NCH larval fish had developed normally with an opened mouth except they could not take food after complete utilization of their egg yolk on the 5th day of development. The possible mechanisms for obtaining such inter-order NCH larval fish are discussed. This is the first report indicating that inter-order NCH larval fish can be obtained in spite of their evolutionary divergence. 相似文献
20.
Monoclonal antibodies specific for type 3 protein kinase C recognize distinct domains of protein kinase C and inhibit in vitro functional activity 总被引:12,自引:0,他引:12
K L Leach E A Powers J C McGuire L Dong S C Kiley S Jaken 《The Journal of biological chemistry》1988,263(26):13223-13230
Monoclonal antibodies (mAbs) which distinguish Type 3 protein kinase C (PKC) from Types 1 and 2 have been obtained from mice immunized with purified Type 3 PKC from rabbit brain cytosol. Most of these mAbs (seven out of eight) selectively recognize Type 3 versus Types 1 and 2 PKC in both enzyme-linked immunosorbent and immunoblot assays. Trypsin treatment of Type 3 PKC reduced the immunoreactivity with 82-kDa PKC and generated immunoreactive fragments of 45 and 35 kDa. The mAbs can be divided into two classes based on their ability to recognize the 45-kDa catalytic fragment (5/8) or the 35 kDa regulatory domain fragment (3/8). Each of the mAbs inhibits phosphorylation of histone or lipocortin by PKC, although the extent of the inhibition varied. Only those mAbs that recognize the 35-kDa regulatory domain inhibited phorbol ester binding. The inhibition of both kinase and binding activities by this group of mAbs was sensitive to the concentration of phospholipid used in the assay. This functional inhibition suggests that these mAbs may be useful for defining the phospholipid binding domain(s) of Type 3 PKC. The mAbs recognized 82-kDa PKC in a variety of cell types; the presence of smaller molecular weight fragments was not consistently found. Distinct immunofluorescence staining patterns were observed with mAbs directed toward different epitopes, suggesting that there may be heterogeneity in the subcellular localization of PKC. The type specificity of these mAbs will make them valuable tools for studying activation and regulation of Type 3 PKC in cell culture model systems. 相似文献