β-actin promotor was cloned from chicken B lympha DT40 cells and used to replace the SV40 promotor of pCDNA3. 1( + )vector,so as to construct the Neomycin resistance express cassette driven byβ 2actin promotor. Then about 2 kb DNA sequenceson each side of the locusof sIgMλgene were inserted into the MCSof the constructed cassette,act2 ing as the homologous arm. PCR identification,restriction enzyme digestion and sequence analysis confirmed that the tar2 geting replacement vector pCDNA2act2neO2HR for sIgMλgene was successfully constructed. This laid the foundation for establishing the model cell lack of sIgMλgene,and further investigating the roles of sIgMλlight chain in the infection of IBDV to DT40 cells.
YoU Lei ming
,
LUo Jun
,
WANG Ai ping
,
ZHANG Gai ping
,
BU Dan
,
Guo Yan an
,
QI Yan hua
. Construction of Knockout Vector for sIgM λ Light Chain Gene from DT40 Cells[J]. Acta Agriculturae Boreali-Sinica, 2009
, 24(3)
: 1
-6
.
DOI: 10.7668/hbnxb.2009.03.001
[1] Cosgrove A S.An apparently new disease of chicken-a-vian nephrosis[J].Avian Dis,1962,6:385-389.
[2] Hirai K,Calnek B W.In vitro replication of infectious bursal disease virus in established lymphoid cell lines and chicken B lymphocytes[J].Infect Immun,1979,25(3):964-970.
[3] Lin T W,Lo C W,Lai S Y,et al.Chicken heat shock protein 90 is a component of the putative cellular receptor complex of infectious bursal disease virus[J].J Virol,2007,81(16):8730-8741.
[4] Nieper H,Muller H.Susceptibillity of chicken lymphoid cells to infectious bursal disease virus does not correlate with the presence of specific binding sites[J].J Gen Virol,1996,77:1229-1237.
[5] Ogawa M,Yamaguchi T,Sefiyono A,et al.Some characteris-ties of a cenular receptor for virulent infectious bursal disease virus by using flow cytometry[J].Arch Virol,1998,143(12):2327-2341.
[6] Yamaguchi T,Iwata K,Kobayashi M,et al.Epitope mapping of capsid proteins VP2 and VP3 of IBDV[J].Archives of Vi-rology,1996,141:1493-1507.
[7] Bergelson J M,Cunningham J A,Droguett G,et al.Isolation of a common receptor for Coxsaekie B viruses and adenovirus-es 2 and 5[J].Science,1997,275(5304):1320-1323.
[8] Berger E A,Nurphy P M,Farber J M.Chemokine receptors as HIV-1 coreceptors:roles in viral entry,tropism,and disease[J].Annu Rev Immund,1999,7:657-700.
[9] Terasaki K,Hirayama H,Kasanga C J,et al.Chicken B lym-phoma DT40 cells as a useful tool for in vitro analysis of pathogeme infectious bursal disease virus[J].J Vet Med Sci,2008,70(4):407-410.
[10] Thomas K R,Capecehi M R.site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells[J].Cell,1987,51(3):503-512.
[11] 李若楠,张彦.转基因动物技术及其应用[J].华北农学报,2007,22:181-184.
[12] 徐永华,常万存,丛笑情,等.动物细胞工程[M].第1版.北京:化学工业出版社,2003:96-97.
[13] 杨晓,黄培堂,黄翠芬,等.基因打靶技术[M].第1版.北京:科学出版社,2003:19-20.
[14] Thomas K R,Deng C,Capecchi M R.High-fidelity gene tar-geting in embryonic stem cells by using!sequence replace-ment vectors[J].Mol Cell Biol,1992,12(7):2919-2923.
