Stephanie bettingen germany

Open in new tab Download slide Construction of plastid transformation vectors for the expression of a chimeric nptII gene under the control of a mitochondrial promoter. Physical maps of the targeting region in the tobacco plastid genome and the two plastid transformation vectors, pSR7 and pSR8, are shown.

Genes above the lines are transcribed from the left to the right, genes below the lines are transcribed in the opposite direction. Sites lost due to ligation to heterologous ends are shown in parentheses. Note that the selectable marker gene for chloroplast transformation, aadA , is under the control of plastid-specific expression signals, whereas the kanamycin resistance gene nptII is driven by mitochondrial expression signals.

With this chimeric mitochondrial-type nptII cassette, two plastid transformation vectors were constructed. In vector pSR7, the nptII has the same transcriptional orientation as the upstream trnfM gene and the downstream selectable marker gene aadA conferring resistance to the aminoglycoside antibiotics spectinomycin and streptomycin; Figure 2. As with this construct, nptII mRNA accumulation could, at least in part, come from co-transcription with trnfM due to read-through caused by incomplete transcription termination; 36 , we also generated a plastid transformation vector with the opposite orientation of the nptII cassette pSR8; Figure 2 , thereby eliminating the possibility of read-through transcription.

The transformation vectors were introduced into plastids on the surface of 0. For both constructs, numerous antibiotic-resistant lines were obtained and successful chloroplast transformation was tentatively confirmed by double resistance tests on medium containing both spectinomycin and streptomycin, a test which eliminates spontaneous spectinomycin-resistant lines 18 , 21 , Putative plastid transformants were passed through additional rounds of selection and regeneration to enrich the transgenic plastid genome and eliminate residual wild-type genome copies 21 , After the third-regeneration round, RFLP analyses were conducted to verify successful plastid transformation, confirm correct transgene integration into the intergenic spacer region between the trnfM and trnG genes Figure 2 and test for the presence of a homogeneous population of transgenic plastid genomes homoplasmy.

Figure 3. Open in new tab Download slide Molecular and genetic analysis of plastid transformants. Fragment sizes for the wild type Wt and the transplastomic lines are indicated in kb. M: molecular weight marker. Note that a very faint wild-type-like band is seen in all transgenic lines. This weak hybridization signal comes from promiscuous DNA that presumably resides in the nuclear genome see text for details.

B Example of a seed assay to confirm homoplasmy of transplastomic lines. Due to maternal inheritance of plastids in tobacco, homoplasmic transplastomic lines produce progeny that is homogeneously resistant to spectinomycin. Lack of phenotypic segregation in the T 1 generation, therefore, demonstrates homoplasmy. Wt: wild-type control displaying spectinomycin sensitivity. In addition to a strong band for the expected restriction fragment, RFLP analysis of both the pSR7 and pSR8-derived transplastomic tobacco lines also showed a faint hybridization signal that corresponds in size to the wild-type fragment Figure 3 A.

Persistence of a wild-type-like hybridization signal even after multiple rounds of selection and regeneration is often seen in plastid transformation experiments and normally, does not come from true heteroplasmy of the plastid transformants. During evolution, large fragments of chloroplast DNA have integrated into the nuclear and mitochondrial genomes 3 and this non-functional promiscuous DNA can produce wild-type-like bands in Southern blot analyses of otherwise homoplasmic transplastomic lines 36 , Seed assays provide a simple and reliable method to distinguish between promiscuous DNA and heteroplasmy When we tested our pSR7 and pSR8-derived transplastomic lines for segregation of the spectinomycin resistance in the T1 generation, the progeny turned out to be homogeneously resistant to the antibiotic, demonstrating homoplasmy of the transgenic plastid genome Figure 3 B, data not shown and confirming our earlier finding that tobacco harbors promiscuous DNA homologous to the targeting region in the plastid genome used in this study 38 , Expression of the mitochondrial-type nptII transgene in plastids We next wanted to test whether the mitochondrial nptII transgene is expressed from the transgenic plastid genome.

RNA accumulation was compared with two transgenic lines that strongly express nptII gene versions under different expression signals: a nuclear-transgenic line containing an nptII transgene under the control of CaMV35S promoter and terminator 39 , 40 and a chloroplast-transformed line in which the nptII was driven by the strongest available plastid promoter, the ribosomal RNA operon promoter P rrn 21 , Read-through transcription due to incomplete transcription termination is quite common in plastids 38 , In fact, a read-through transcript with the downstream aadA has been observed before for other transgenes inserted into the plastid transformation vector used to construct pSR7 [vector pRB94, 19 , 38 ].

Figure 4. Open in new tab Download slide Expression of the mitochondrial-type nptII transgene in tobacco plastids. A dilution series for a chloroplast transformant P rrn : nptII , in which nptII is driven by the strongest known plastid promoter, the ribosomal RNA operon promoter P rrn , is also shown.

The amount of mRNA loaded in each lane is indicated, the sizes of the bands of the molecular weight marker are given at the left. Additional, minor RNA species were not further characterized. The amounts of total soluble protein TSP loaded in each lane are indicated. Compared to the strongly expressing nuclear and plastid nptII gene versions, the sensitivity of the anti-NptII antibody is insufficient to clearly detect the protein expressed from the mitochondrial-type nptII in plastids see Figure 5.

Next, we analysed accumulation of the nptII gene product, the neomycin phosphotransferase protein, NptII. Using a specific anti-NptII antibody, protein accumulation in SR7 and SR8 lines was compared with that in the two control transformants. As NptII protein levels in the SR7 and SR8 transformants were on the borderline of detectability, we conducted kanamycin resistance assays to ultimately confirm expression of the mitochondrial-type nptII gene in plastids at the protein level and, simultaneously, test for the accumulation of active NptII enzyme.

Resistance tests were performed both with leaf explants exposed to regeneration media containing different concentrations of kanamycin Figure 5 A, data not shown and with seeds germinated in the presence of the antibiotic Figure 5 B. Both sets of tests revealed a significant resistance to kanamycin in the SR7 and SR8 transplastomic lines Figure 5 , confirming expression of the nptII from the mitochondrial expression cassette at the protein level and moreover, demonstrating that the NptII protein produced from the mitochondrial-type gene in plastids is enzymatically active.

Interestingly, in both assays, kanamycin resistance was significantly stronger in the SR7 transplastomic lines than in the SR8 lines Figure 5. Figure 5. Open in new tab Download slide Confirmation of the expression in plastids of functional enzyme from the mitochondrial-type nptII by phenotypic assays. A Determination of kanamycin resistance by exposure of leaf explants to kanamycin-containing plant regeneration medium.

After 35 days, only regeneration from leaf pieces of the control plastid transformant line P rrn : nptII containing an nptII under the control of the strongest available plastid promoter is seen upper picture. However, after 70 days on kanamycin-containing medium, regeneration of SR7 and SR8 lines is obtained, whereas explants from the wild-type control Wt and a control plastid transformant harboring just the selectable marker gene aadA P rrn : aadA are fully bleached out due to their sensitivity to the antibiotic.

B Analysis of kanamycin resistance by seed assays. Note that in both A and B , the higher level of nptII expression in the SR7 lines due to the presence of additional read-through transcripts; see Figure 4 correlated with stronger phenotypic resistance to kanamycin. Analysis of transcriptional activity and mapping of transcription start sites The data described above imply transcription of the plastome-integrated nptII -cassette under control of the mitochondrial atpA promoter.

However, the observed monocistronic nptII transcript Figure 4 A could come from transcription initiation at the P atpA promoter or, alternatively, from processing of larger precursor RNAs whose transcription initiates at an upstream plastid promoter.

Figure 6. Amplified products were separated by agarose gel electrophoresis, fragment sizes of the DNA molecular weight marker are given in base pairs. Below the chromatogram, the sequence of the atpA promoter region with the core promoter motif marked in boldface letters is shown. The transcription initiation site is underlined and marked by the bent arrow.

This hybridization experiment confirmed the active transcription of the nptII gene in the transplastomic lines SR7 and SR8. As expected, run-on transcripts from wild-type chloroplasts used as control did not hybridize to the nptII probe on the filter. Remarkably, the hybridization signals for the nptII transcript were distinctly stronger than the signals for the resident chloroplast genes rpoB and clpP , indicating that the mitochondrial promoter was more efficient in driving plastid transcription than the investigated chloroplast rpoB and clpP NEP promoters.

Figure 7. Open in new tab Download slide Detection of newly synthesized nptII transcripts in isolated transgenic plastids. The spotting scheme and dot-blot autoradiograms of chloroplast run-on transcription experiments are shown. Transcriptional activity from the mitochondrial P atpA promoter driving the chimeric nptII gene in plastids is compared with that of representative resident plastid genes rpoB, clpP, psbA.

Transcription from the mitochondrial atpA promoter initiated at the identical site in mitochondria, in transgenic chloroplasts and in an in vitro transcription system using purified organellar RNA polymerases. It is well established that both prokaryotic and eukaryotic RNA polymerases require transcription factors for promoter recognition.

In contrast, promoter recognition by the bacteriophage T7 RNA polymerase is an intrinsic property of this single-polypeptide enzyme This remarkable feature may have been retained during the evolution of organellar phage-type RNA polymerases from an ancestral bacteriophage enzyme 15 , However, in vivo , effective transcription by the yeast enzyme is known to require transcription factors A similar situation has been suggested for transcription in Arabidopsis mitochondria and chloroplasts, since RpoTm and RpoTp were not able to recognize in vitro all of the promoters they utilize in vivo , and RpoTmp did not even efficiently recognize any promoter in vitro In the present investigation, we also observed no specific recognition of P atpA by RpoTmp in in vitro assays.

These findings raise interesting evolutionary considerations. First, the data from in vitro transcription assays support the idea that RpoTm and RpoTp have kept the intrinsic capability for promoter recognition during evolution from an ancestral phage polymerase, while RpoTmp has lost this property The plastid enzyme RpoTp was less efficient than the mitochondrial RNA polymerase in correct initiation from the mitochondrial atpA promoter in vitro , possibly reflecting an evolutionary adaptation of RpoTp to plastid promoters.

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But, after Oct. On that day, the gymnasium within its walls would bear Price's name. Those individuals who lobbied for the change already knew the joy and pain conveyed behind the wrinkles on Price's face. Those 40 or so individuals who attended the ceremony became familiar with, or were already exposed to, his decades of dedication to the Eifel community.

But what about those who may not know the man for whom the gymnasium is named? The answer lies in learning more about the lives of those whom he's touched throughout the decades of his military and civilian service as well as from the man himself. Pictures of memories spanning decades of his service to the local German community and the U.

Air Force adorn the four walls of his office at the 52nd Force Support Squadron building. And Price, sitting at his desk as the 52nd FSS special events coordinator, reaches over to answer a ringing phone. Hey, Big Man. Those accustomed to conversation with Price have no doubt heard him use that catchphrase, one that started when he got involved in the world of basketball.

Born and raised in a small town in Pennsylvania, Price joined the U. Air Force in I matured, learned to lead and learned how to be happy. He said his desire to be competitive at anything he did showed during this time. And his commitment to the Air Force Core Values carried him on to earn the same title three more consecutive times after that.

When he hung up his service dress coat for the last time, his ribbon rack sported the following medals: a Bronze Star, a Purple Heart, a Meritorious Service Medal, an Air Force Commendation Medal and other numerous campaign awards.

But that door closing didn't signal his final round of service for Airmen. And while he said that he felt unprepared for civilian life, a certain wingman would help him out. Frederick Kyler, was a very good friend of mine," he said. It cost 35 cents a sack for the first five sacks.

I was making so much money that I was afraid they were going to arrest me! He said he noticed a few occasions where children arrived at the center without anything to eat. Despite not having a food handler's card, Price served them breakfast, in line with the first sergeant's creed of putting people first.

Price's transition to working with youths soon led him to coaching, and he soon took over as the athletic director for Bitburg Air Base. Although with no previous experience, his gung-ho spirit and determination inspired him to make his program the best in Europe. He was a high school teacher and the smartest basketball man I ever knew.

But he couldn't lead a goat to get a drink; I could get the goat to drink. So I used his brains to teach, and we took off and won time after time through his knowledge and my ability to scream and shout. That's how we got by. I always carried 12 guys and only dressed For the other two guys, one would get the bus, and the other would pick up the dirty towels.

Air Forces in Europe and Air Forces Africa major command history, like his Air Force career, speaks for itself for those on the bleachers or on the benches: Eleven continental sport conference championships. Air Forces Central Command tournaments. Back-to-back victories in the and "Basketball Tip-off Classics. There can only be one "winningest.

But as I look back on it now and some of the friends I made, some of my ball players - it was just out of this world, and I was fortunate to be a part of it. Many may know Price's devotion to both Airmen in uniform and athletes on a court. Yet deeper than all of the victories or trophies, he said his family always comes first. As a young staff sergeant, Price said he fell in love with a young German woman named Hildi, who worked at the community center snack bar at Bitburg Air Base.

Fifty-six years later, Mr. Price remain inseparable, and Price even said he'd marry her all over again if given the chance. The newlywed Prices soon doubled their family with the births of their daughter Bee Bee and son Robin. It was a great childhood. If you failed or succeeded, he was OK with it. But if he realized that you didn't do your best Kader added that growing up in the presence of a champion proved to be both great and challenging. Once I became a teenager, it wasn't easy.

A lot of the Air Force guys wouldn't date me because my dad was George Price. But those last two developments took a backseat to a closer and more personal tragedy within the Price household as Robin received a diagnosis with cancer. I didn't do it no more. Over the years, the position brought him near the same courtside action he had been familiar with for so many years, yet he said something still felt missing. I just had no energy to develop it. So I just let it go, and I haven't coached since.

Of this area, 0. Of the rest of the land, 0. Out of the forested land, Of the agricultural land, Bettingen is split into two settlements. Dorf Bettingen is an Alemannic settlement, located in a small basin. Chrischona pilgrimage centre established on one of the hills. As early as a church was built on the heights, dedicated to Saint Chrischona.

Since there have been a hospital and retirement homes in the area. A landmark for Bettingen, the Swisscom-Sendeturm St. Coat of arms[ edit ] Demographics[ edit ] Bettingen has a population as of July [update] of 1, It has changed at a rate of 2.

There were or Ignoring immigration and emigration, the population of Swiss citizens decreased by 5 while the foreign population increased by 5. There were 4 Swiss men who immigrated back to Switzerland. At the same time, there were 7 non-Swiss men and 11 non-Swiss women who immigrated from another country to Switzerland.

The total Swiss population change in from all sources, including moves across municipal borders was a decrease of 14 and the non-Swiss population increased by 21 people. This represents a population growth rate of 0. There were married individuals, 61 widows or widowers and 55 individuals who are divorced.

Out of a total of households that answered this question, Of the rest of the households, there are married couples without children, married couples with children There were 18 single parents with a child or children. There were 4 households that were made up of unrelated people and 19 households that were made up of some sort of institution or another collective housing.