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Investigations on a clinically and functionally unusual and novel germline p53 mutation.

Rutherford J, Chu CE, Duddy PM, Charlton RS, Chumas P, Taylor GR, Lu X, Barnes DM, Camplejohn RS - Br. J. Cancer (2002)

Bottom Line: Surprisingly two assays of p53 function gave apparently wild-type results on peripheral blood lymphocytes from this individual.These results led us to carry out more detailed functional tests on the mutant protein.However, surprisingly, data from irradiated peripheral blood lymphocytes and transfected Saos-2 cells, suggested that this truncated, mutant protein retains significant ability to induce apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Richard Dimbleby Department Cancer Research, Guy's, King's and St Thomas' School of Medicine, St Thomas' Hospital, London SE1 7EH, UK.

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Western blot. The 7 base pair mutation was cloned into the mammalian expression plasmid pC53-SN3, which was transfected into Saos-2 cells. A Western blot was carried out using lysates from these cells, cells transfected with WT p53 and PHA stimulated lymphocytes from the patient. The blot showed WT p53 with the expected size of 53 kDa (lane 1) and the p53 from the patient's lymphocytes also at size 53 kDa (lane 2). The cells transfected with the manufactured mutation showed a protein on the blot at about size 27 kDa (lane 3).
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fig4: Western blot. The 7 base pair mutation was cloned into the mammalian expression plasmid pC53-SN3, which was transfected into Saos-2 cells. A Western blot was carried out using lysates from these cells, cells transfected with WT p53 and PHA stimulated lymphocytes from the patient. The blot showed WT p53 with the expected size of 53 kDa (lane 1) and the p53 from the patient's lymphocytes also at size 53 kDa (lane 2). The cells transfected with the manufactured mutation showed a protein on the blot at about size 27 kDa (lane 3).

Mentions: The manufactured 7 base pair insertion mutation was cloned into a mammalian expression plasmid and was transfected into Saos-2 cells. Cell lysates were made of these cells and of Saos-2 cells transfected with wild-type p53. PHA stimulated lymphocytes from the patient were also lysed and a Western blot was run using all three lysates (Figure 4Figure 4


Investigations on a clinically and functionally unusual and novel germline p53 mutation.

Rutherford J, Chu CE, Duddy PM, Charlton RS, Chumas P, Taylor GR, Lu X, Barnes DM, Camplejohn RS - Br. J. Cancer (2002)

Western blot. The 7 base pair mutation was cloned into the mammalian expression plasmid pC53-SN3, which was transfected into Saos-2 cells. A Western blot was carried out using lysates from these cells, cells transfected with WT p53 and PHA stimulated lymphocytes from the patient. The blot showed WT p53 with the expected size of 53 kDa (lane 1) and the p53 from the patient's lymphocytes also at size 53 kDa (lane 2). The cells transfected with the manufactured mutation showed a protein on the blot at about size 27 kDa (lane 3).
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2746598&req=5

fig4: Western blot. The 7 base pair mutation was cloned into the mammalian expression plasmid pC53-SN3, which was transfected into Saos-2 cells. A Western blot was carried out using lysates from these cells, cells transfected with WT p53 and PHA stimulated lymphocytes from the patient. The blot showed WT p53 with the expected size of 53 kDa (lane 1) and the p53 from the patient's lymphocytes also at size 53 kDa (lane 2). The cells transfected with the manufactured mutation showed a protein on the blot at about size 27 kDa (lane 3).
Mentions: The manufactured 7 base pair insertion mutation was cloned into a mammalian expression plasmid and was transfected into Saos-2 cells. Cell lysates were made of these cells and of Saos-2 cells transfected with wild-type p53. PHA stimulated lymphocytes from the patient were also lysed and a Western blot was run using all three lysates (Figure 4Figure 4

Bottom Line: Surprisingly two assays of p53 function gave apparently wild-type results on peripheral blood lymphocytes from this individual.These results led us to carry out more detailed functional tests on the mutant protein.However, surprisingly, data from irradiated peripheral blood lymphocytes and transfected Saos-2 cells, suggested that this truncated, mutant protein retains significant ability to induce apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Richard Dimbleby Department Cancer Research, Guy's, King's and St Thomas' School of Medicine, St Thomas' Hospital, London SE1 7EH, UK.

Show MeSH
Related in: MedlinePlus