Cell-specific promoter in adenovirus vector for transgenic expression of SERCA1 ATPase in cardiac myocytes.
Cell-specific promoter in adenovirus vector for transgenic expression of SERCA1 ATPase in cardiac myocytes.
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腺病毒载体中的细胞特异性启动子，用于心肌细胞中 SERCA1 ATP 酶的转基因表达。
DOI：
10.1152/ajpcell.1998.274.3.c645
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Ordahl,CP
中科院分区：
文献类型：
--
作者：
Inesi,G;Lewis,D;Sumbilla,C;Nandi,A;Strock,C;Huff,KW;Rogers,TB;Johns,DC;Kessler,PD;Ordahl,CP
腺病毒介导的鸡骨骼肌肉瘤（内质网）Ca2+- atp酶（SERCA1） cDNA的转移，在细胞特异性心肌肌钙蛋白T （cTnT）启动子片段（- 268碱基对）的控制下，在培养的鸡胚胎心肌细胞中选择性表达，或在组成型病毒[巨细胞病毒（CMV）]启动子的控制下，在肌细胞和成纤维细胞中非选择性表达。在最佳条件下，几乎所有培养的心肌细胞都表达转基因SERCA1 atp酶。表达以细胞膜为目标，并在亚细胞组分中恢复，其模式与内源性SERCA2a atp酶相同。与对照肌细胞相比，转基因SERCA1表达增加了细胞匀浆中atp依赖性（和thapsigarin敏感）Ca2+转运活性的四倍。虽然CMV启动子比cTnT启动子更有活性，但通过调节感染介质中腺病毒斑块形成单位滴度，在两种启动子的控制下，功能酶的转基因表达达到了上限。在任一启动子的控制下，SERCA1转基因表达对整个肌细胞的细胞质Ca2+浓度瞬态和张力发展也有类似的影响。我们的实验表明，重组腺病毒载体中的细胞特异性蛋白启动子可在心肌细胞中高效、选择性地表达一种膜结合的功能性酶。
Adenovirus-mediated transfer of cDNA encoding the chicken skeletal muscle sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA1) yielded selective expression in cultured chick embryo cardiac myocytes under control of a segment (−268 base pair) of the cell-specific cardiac troponin T (cTnT) promoter or nonselective expression in myocytes and fibroblasts under control of a constitutive viral [cytomegalovirus (CMV)] promoter. Under optimal conditions nearly all cardiac myocytes in culture were shown to express transgenic SERCA1 ATPase. Expression was targeted to intracellular membranes and was recovered in subcellular fractions with a pattern identical to that of the endogenous SERCA2a ATPase. Relative to control myocytes, transgenic SERCA1 expression increased up to four times the rates of ATP-dependent (and thapsigargin-sensitive) Ca2+transport activity of cell homogenates. Although the CMV promoter was more active than the cTnT promoter, an upper limit for transgenic expression of functional enzyme was reached under control of either promoter by adjustment of the adenovirus plaque-forming unit titer of infection media. Cytosolic Ca2+concentration transients and tension development of whole myocytes were also influenced to a similar limit by transgenic expression of SERCA1 under control of either promoter. Our experiments demonstrate that a cell-specific protein promoter in recombinant adenovirus vectors yields highly efficient and selective transgene expression of a membrane-bound and functional enzyme in cardiac myocytes.
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影响因子：
20.1
作者：
LOMPRE, AM;LAMBERT, F;SCHWARTZ, K
通讯作者：
SCHWARTZ, K
含有多种细胞类型的原代心脏培养物中转染基因表达的单细胞分析。
影响因子：
2.7
作者：
Antin,PB;Mar,JH;Ordahl,CP
通讯作者：
Ordahl,CP
DOI：
10.1172/jci116577
发表时间：
1993-07
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
L. Kirshenbaum;W. Maclellan;W. Mazur;B. A. French;M. Schneider
通讯作者：
L. Kirshenbaum;W. Maclellan;W. Mazur;B. A. French;M. Schneider
DOI：
10.1073/pnas.90.24.11498
发表时间：
1993-12-15
影响因子：
11.1
作者：
KASSEISLER, A;FALCKPEDERSEN, E;LEINWAND, LA
通讯作者：
LEINWAND, LA
DOI：
10.1073/pnas.52.6.1456
发表时间：
1964
影响因子：
11.1
作者：
M. E. Carsten
通讯作者：
M. E. Carsten