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·38 · Prog.Biochem.Biophys. 1999;26 (1)
Construction, Expression and Purification of
Calponin T184A:a Mutant of Calponin.CAO
1 , , (Tang D C, Xiang J Z, Lu W T). Sheng, TANG Da-Chun, LUO Jian-Yun (Key
Calponin.
(Prog Biochem Biophys), 1996, 23 (4):32 ～329 Laboratory of Pulmonary Disease, Health Ministry
2 Tang D C, Kang H M, Jin J P, et al.Structure-function relations of China;Department of Pathophysiology , Tongj i
of calponin.Biophys J, 199 , 68 (2Pt2):A163
3 , , (Cao S, Tang D C, Luo J Y). Medical University , Wuhan 430030, China).
. (Acta Abstract Calponin cDNA was site-mutated (Thr184
Universitatis Medicinal Tongji), 1997, 26 (1):67 ～68
4 J, E F, T (Sambrook J, Fritsch E ※Ala184 )by overlap extension PCR method to
F, Maniatis T )., . (Molecu- reduce the opportunity for calponin to be phosphory-
lar cloning:a laboratory manual). (Beijing):
(Science Press), 1992.23 ～371 lated by PKC or CaM K Ⅱ.The PCR product was
Winder S J, Walsh M P.Calponin:thin filament-linked regulation sequenced.The mutated cDNA w as then recombined
of smooth muscle contraction.Cell Signal, 1993, 5 (6):677 ～
into an expression vector pAED and transferred into,
4
682
6 Sobue K, Sellers J R.Caldesmon:a novel regulatory protein in expressed in .coli BD (DE )induced by IPTG.
21 3
smooth muscle and nonmuscle actomyosin systems.J Biol Chem,
1991, 266 (19):1211 ～ 12118 The expression level was up to 1 %of total bacterial
7 Tang D C, Stull J T,
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