Phytase expression in transgenic soybeans: stable transformation with a vector-less construct

被引：33

作者：

Gao, Xiao Rong ^{[1
]}

Wang, Guo Kun ^{[1
]}

Su, Qiao ^{[1
]}

Wang, Yan ^{[1
]}

An, Li Jia ^{[1
]}

机构：

[1] Dalian Univ Technol, Dept Biosci & Biotechnol, Dalian 116024, Peoples R China

来源：

BIOTECHNOLOGY LETTERS | 2007年 / 29卷 / 11期

关键词：

pollen tube pathway transformation; linear DNA; phytase gene; transgenic soybean;

D O I：

10.1007/s10529-007-9439-x

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A minimal linear gene cassette (35S-phytase gene-nos) with T-DNA borders was acquired by PCR and directly introduced into soybean through the pollen tube pathway. A total of 13% of T-1 plants were positive for phyA by specific PCR. Southern blot analyses showed that phyA insertions were harbored stably in T-2 progeny. Phytase expression level increased 2.5-fold over a 6-week period; its highest activity was 150 U/mg protein, compared to 56 U/mg protein in untransformed controls. Activity of phytase increased to 125 FTU/kg in T-3 transgenic seeds as compared to 64 FTU/kg in wild-type plants.

引用

页码：1781 / 1787

页数：7

共 24 条

[1]

Carlson J. B., 1987, SOYBEANS IMPROVEMENT, P95

[2] Factors influencing regeneration of soybean from mature and immature cotyledons [J].

Franklin, G ;

Carpenter, L ;

Davis, E ;

Reddy, CS ;

Al-Abed, D ;

Abou Alaiwi, W ;

Parani, M ;

Smith, B ;

Goldman, SL ;

Sairam, RV .

PLANT GROWTH REGULATION, 2004, 43 (01) :73-79

[3] Linear transgene constructs lacking vector backbone sequences generate low-copy-number transgenic plants with simple integration patterns [J].

Fu, XD ;

Duc, LT ;

Fontana, S ;

Bong, BB ;

Tinjuangjun, P ;

Sudhakar, D ;

Twyman, RM ;

Christou, P ;

Kohli, A .

TRANSGENIC RESEARCH, 2000, 9 (01) :11-19

[4]

GONG ZZ, 1988, SCI SINICA B, V9, P1080

[5]

HAMADA A, 2004, PLANT BIOTECHNOL J, V2, P1

[6] A novel phytase with sequence similarity to purple acid phosphatases is expressed in cotyledons of germinating soybean seedlings [J].

Hegeman, CE ;

Grabau, EA .

PLANT PHYSIOLOGY, 2001, 126 (04) :1598-1608

[7] Integration of T-DNA binary vector 'backbone' sequences into the tobacco genome: Evidence for multiple complex patterns of integration [J].

Kononov, ME ;

Bassuner, B ;

Gelvin, SB .

PLANT JOURNAL, 1997, 11 (05) :945-957

[8] Secretion of active recombinant phytase from soybean cell-suspension cultures [J].

Li, J ;

Hegeman, CE ;

Hanlon, RW ;

Lacy, GH ;

Denbow, DM ;

Grabau, EA .

PLANT PHYSIOLOGY, 1997, 114 (03) :1103-1111

[9] Linear transgene constructs lacking vector backbone sequences generate transgenic rice plants which accumulate higher levels of proteins conferring insect resistance [J].

Thi Loc N. ;

Tinjuangjun P. ;

Gatehouse A.M.R. ;

Christou P. ;

Gatehouse J.A. .

Molecular Breeding, 2002, 9 (04) :231-244

[10]

Luo Z., 1989, PLANT MOL BIOL REP, V7, P69, DOI [10.1007/BF02669248, DOI 10.1007/BF02669248, DOI 10.1007/BF00269517]

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