- n c CONG N G H $ SINH HOC TOAN QUOC 2013
S A N G L O C - D O T B I ^ N G E N G A Y BENH C A N G DA Nguyen ThCiy Dirong \ Manfred Wetinert \ Nong Vdn Hai *
* W#n Nghi§n aiu he gen, W$n Harj Idm Khoa ftpc v^ C6ng ngh^ Vi§t Nam Vi$n AfgfiiAj ciru di truyin ngiriri. Gre'ifswald, C^ng hda UBn bang Buc
T6M TAT
B ^ cSng da (Restrictive dCTmopalhy- RD) li b ^ h di Iniyk lan r ^ hi&n gSp, do dOt bi£n trong gen ZMPSTE24 ho5c LMNA gay ngn. Tre m^c b$nh n ^ c6 btSu hi$n 13m s ^ g nhu da c ^ g mong dl i^di nliin thiy ca oifich mAu dudi da vh dhu ctik trong tuIn dm sau sinh. Trong nghifin cthi oky, chiing t6i ti^ hinh sing IQC tim dOt bign tiong hai gen ZMPSTE24 hojc IMNA cua mOt gia dinh b§nh nhdn cSng da. Sur dung k^ thu$t PCR va cdc c$p moi dupc tliiet kS dac h i ^ cac doan exon ciia ck hai grai trdn da dupc nhdn thinh cfing. T i ^ do, chiing t6i dS tiin hinh dpc trinh tu gen dye ti^ san pham PCR tinh sgch. Sau khi so sinh trinh Vif gen thu dupc vdi cic trinh tv gen c6ng b6 tr&i Genbank {NM_005857 ciia ZMPSTE24 vi NM_170707 ciiaiAflWX h$nh nhSn mang hai dftt biin d) hpp tu trong gen ZMPSTE24, c.SOdelA vi c.1085 -1086in3T. EKH biin c.50delA di truyen tit b6, Idiong tim thSy 6 386 allele oia quan the n§u6i binh thuimg. DOt biin cdn l^i di truyin tCr m? li dOt biio fliudng gap 6 b§Dh nhfin cing da. 6 gen LMNA, khdng tim th4y d$t biSn nio. Kit qui nghiSn cihi niy them m^t ddng gdp nghiSn curu bfnh di truyin ti mirc dO phfin tCr.
Tir khda: B ^ cang da, dOt b i ^ LMNA, PCR, ZMPSTE24
nCfoAu
B$nh Gang da (RD) Id b$nh di truySn bim sinh hiem g$p. RD \d b^nh OfC ky nguy hilm, gay c h ^ ngydri chi m$t trong hai tuSn diu sau sinh. Thai nhi thu'd'ng cd cdc dku hi$u nhu- ch|m phdt triln vd hlnh ihdi dj dang. Khi du'Q'c sinh ra, b§nh nhdn cd mpt s6 tri^u chirng idm sang nhu: da rat cing, khdng cd kha ndng dan h6i, nui d* cdc khdp cu- d$ng, mi^ng nhd ludn mcr hlnh chO 'O', mui nhpn nhd, ldng mi thu-a ho$c khdng cd. B$nh nhdn chet sd-m ^ u ^ g la do bj suy hd hap n$ng (Navarro ef al., 2004). DOt bi$n trong cd hai gen ZMPSTE24 vd LMNA diu gdy b§nh RD. Trong dd, ddt biln trong gen ZMPSTE24 Id nguyen nhdn gdy b$nh cua da s6 b$nh nhdn RD (Moulson ef al., 2005).
Gen ZMPSTE24 m§ hda protein metalloprotease STE24 (ZMPSRE24) nam trdn nhiim sSc t h i s6 1 {1p34) Gen ndy cd kich thu^c khoang 36 kb vd gom 10 exon. ZMPSTE24 tham gia vdo hai trong tion bir6c xi> l<^ sau djch ma cCia tiln LMNA (prelamin A) (Hlnh 1) (HoHz etal., 1989, Sinensky etal., 1994). Gen LMNA ma hda cho lamin Avd lamin C ndm trdn nhilm sic t h i s6 1 (1q22). Gen ndy cd kfch lliudddioang 57 kb vd g6m 12 exon. Lamin Avd Old cdc protein cau tnic nam ngay du'di mdng nhdn. ChOng tham gia vdo mOt s6 hogt ddng chfnh cCia t l bdo nhur dilu hda bilu hi§n gen^
sao chdp DNA vd si>a chO^ sai hdng (Prdtocimer ef al., 2009). Tnjng khi lamin C dirgrc tgo thdnh ngay sau djch ma da cd vai trd cua mpt protein truing thdnh thi lamin A phii.bai qua qua trinh biln dSi sau djch md tgo ra lamin A budng thdnh cd chi>c ndng tCr tiln lamin A.
DOt biln trong gen ZMPSTE24 diu tien du-grc xdc djnh d> cdc b^nh nhdn logn co thdi dirang hdm dgng B (It^andibuloacral Dysplasia- MAD-B) (Aganwai ef al.. 2003), sau dd dygc tim thiy it b^nh nhdn cdng da (Moulson ef a/., 2005, Navarro ef a/., 2005, Navarro ef al, 2004). Ngodi ra, dOt bien gen ZMPSTE24 cung du'g^ tim thiy it nhilu b$nh nhdn cdng da khdc (Li 2010) vd d bOnh nhdn gid tnrirc tu6i (Hutdiingon Gilford Progeria s^drome- HOPS) cd cd bilu hign ldm sdng cOa bOnh MAD (Ben Yaou et al., 2011). Trong khi dd, dOt biln gen LMNA Id nguyen nhdn gdy ft nhlt 10 b§nh khdc nhau duvc gpi Id nhdm b§nh lamin vdi mOt so bilu hiOn lam sdng chOng chdo. MOt trong nhOm bdnh nay cDng id bgnh HGPS vdi nguyen nhdn gdy bgnh xuit phdt to- SV tidi lOy dgc t l tiln lamin A.
DOt biln c.1824C>T khdng Idm thay doi amino add Gly608Gly nhu'ng lam thay d l i v\ trf nhgn bilt sau phien ma du'^c tlm thiy d nhilu b^nh nhdn mic bfinh gid tordc tuoi (Eriksson ef al., 2003). BOt biln ndy cung du'vc tim thiy it bOnh nhdn mic b|nh cdng
da (Navarro et ai. 2004). Lamin A trwOTig thinh
Hlnh 1: Qui trinh xi> ly sau djch ma cua tien lamin A (prelamin)
HOl NGHj KHOA HQC C6NG NGH$ SINH HOC l'''-^^'^'^^lS^k^pa\tiam^'^i
Bd, vilt ndy trinh bdy kit qud sdng k?c ddt b^n trong hai gen ZMPSTE24 vd LWWA * /"Ot gia dinh bgnh nhdn mic b$nh cdng da. I^t qua ndy se gdp phin ddng k l trong vigc cung d p thong tin chinh xdc cho chin dpdn bgnh cdng da a g | i dogn sdm gdp ptiln vdo chan dodn b#nh tnrdc sinh.
VAT U^U VA PHl/ONG P H A P
v|tii$u
Mlu mdu cua cd gia dinh cd b$nh nhdn cang da do Bgnh vign Da, COng hda Uen bang Qdc cung d p va dirg'c luu giir i, -80°C tgi Vi$n Nghien cihi di truyin a>u ngudi, Greifswald.
Cdc cdp mli si> dyng d& nhdn cdc dogn exon cfla gen ZMPSTE24 va LMNA diwyc thilt k l d^a tren trinh tif gen da dir<7C cdng b l trtn GenBank vdi md s l NM_005857 cua ZMPSTE24 vd NM_170707 cua LAfAM vd anh hu*ng vj fri cua cdc apt biln- Mli dupc thilt k l d viing intron each exon khoang 150- 250 bp. Ngodi ra, d l friugn tign cho viOc dpc trinh tif frvc tilp ti> san phim PCR, exon 1 cua gen LMNA se duvc nhdn l§n thdnh 2 dogn do nd cd kich thu*c khd ddi.
Trinh t v c i c d p m i i nhfln 12 ao?n exon cua gen LMNA nhu sau: S- CCCAGATCCCGAGGTCCGAC -3', j . AGCCCTGCGTTCTCCGrrrC - y . 5- ACATCGACCGTGTGCGCTC -3'. 5'- CCTCTCCACTCCCCGCCA -3', 5 -
GGATGCCCTCTCCTGGTAAT -S". 5*- GCTCTG AAATCAGGTGACAGG -3', 5'- rrCTTGTGTTCTGTGACCCCTT -y. 5'- CCCAAG TCTGTCATCACCCA -S", S- GCCCCTGGGTCTTGGCCTCC -3', 5'- GGCAGTGAG GGAACCAATCGA -3', 5- CCTCCACCCCTCCCAGTCAC - ? , ff- TGCATCCGGCCCAGA CTCTA -3'. 5'- CAAACCCTCCCACCCCCC -3', S- CCAGTTGCCGGGCCAGAG -3:. ff- CCC CACTTGGTCTCCCTCTCC -3', 5 - CCCTGATGCAGCTGTATCCCC -3', ff.
CCAAGAGCC TGGGTGAGCCTC -3', 5'- GACACTTACCCCAGCGCTCC -3', 5 - CAGGTGGTGACGGTGAGTG -3', S- CAGCTGGCTCCGATGTTG -3". S- ACCCTTCCCTGGCCCTGAC -3-. 5'- CACCTGGGTTCCCTGTTCAAG -3', S- TGGTCA GTCCCAGACTCGCC -T. 5*- CGCCTGCAGGATTTGGAGA -3', 5'- TGAGGGATGGGG GAGATGCT -S", y.
GGGTGGGCATGAGGTGAGGA J ' .
Trtnh t v c i c C^p m&i nhfin 10 doan exon cua gen ZMPSTE24 nhir sau: 5'- GGGACTAGGGAGAGTCTGCAA -3' 5'- GTATGGGGGTCAGGTrGGAC -3'. S- TGGCAAGCTATAAACCATTCG -3'. ff- TGA AAATGAAAACAACCAGACA ^ . 5'- CCGTACTGGCCTCTTTTGTT -S". S- GAAAGC CTGCCAAGCTAAAA -3', 5'- TTGATTTGTTrGCCAGTAGTTCA -3', 5'.
CAGGACAAA AGCACAGAAGTTTT -3", ff- CCAGTTTCTCAGTTTCTTGTGG -3', 5*- TCTCACCAAGGA ACTTTTGC -3' S- GACATTTACTTTTCAGGTTCTTGTCA -S*. ff- TCAAATAAAACA AACCACTTGGA -3'. 5'- CTCCAAAG6ACCCCAAA -3' ff- TTTTGAGTTGTCACA GGAACTG -3', 5"- TGAAGGGCTATTACTGGGTTAAA -3*. 5'- CCTCTCATGCCTGCCATA GT -3' i?- TGATCCCATAGTGAAATCAGCTT -3'. 5"- GATTTGAAGCAGGCAAGAGC -3', S- GGCAGTGGCTAAAACCCTTT -S', S- TGCTGCCAGGACAGAAATAA -3'.
Phiromg phdp Tich chl4t DNA t6ng s6
Dung dfch NaCI 6M dlK?c sir dyng d l tdch chiet DNA tong s l h> mdu ngu*i. Mdu ching dOng frwig EDTA duvc n>a ki d l dung gidi t l bdo hlog d u bing lysis buffer. Dung djch SDS 20% vd proteinase K duvc si> dgng fle phd vd nhdn tl bdo bgch d u vd phdn huy protein. DNA tong s6 duvc phdn tdch nhd b l sung NaCI 6M vd kit tua bang isopropanol.
Nhin gen ZMPSTE24 vi LMNA
Cdc dogn exon cOa hai gen ZMPSTE24 vd LMNA dirgrc khuich dgi bing ky thugt PCR. Mli phan dng (25 u|) bao gim cdc thdnh phin: 0,125^pl T M DNA polymerase (SU); 2.5 pi dOm 10X, 0,5 pi dNTPs (100 mM); 1 ul mli loai mil (10
^ f f f ^ ^ ' ^ ^ o ? ' ! , ' ^ " ^ "^"^ " ' " ' ^ " - ^ ^ 3 P^"^*- 35 chu k* {96"C 20s, 55°C - 64°C 45 gidy, 72''C 45 gidv) 72''C 10 SJ^.I^ ^i ^ ^ • ^ ' ^ K P 1 ^* i^-^" ''"^ '^'^'^ ''^"S ^^ ° ^ " ^ J^'^™ PCR purifiLton cua Hang Thenno baentitic ttwo phirtmg ptiap cua nhd sdn xuit.
Xic dfnh trinh tv gen
A ^ S l ' ^ r ' S ^ ^ t ' Z ^ . ^ ^ ^T-.'^f'"]' ' ' ^ " "^ ""'"S '"*>' ai^i ' ^ " 1 ' ^V <V aOng ABI PRISM 3100 Gaelic mJm * h h J ^ o K ? " " ' ° ^•'-<'°™=' cia ZMPS7E24 va NMJ70707 cOa LMW/l^ng phS«
K ^ QUA VA THAO LUAN
Nhfin edc dofn excHi cua gen ZUPSTE24 vfi LJWN>i DNA t6ng s6 tinh sach ti> mfiu dt/pc si>
gen LMNA thu a V c O n T a cic bang « c ( ! S K , ? ^ ; „ T . h ' ' « ' ' . * l \ ^ . " " * ' " '"'='' '^t*" =*= ' ^ n exon cui
Xic ilinh mnh t v cic doan exon ciJa gen 2MPS7E24 y i LUNA ^ D^ Wiing dinh chfnh xic san oham PCR Id r.^.. * , « • ._ • dmre Unh each d w c liip luc x i c S „ h M h .^ ^ f " " " cua hai gen ZMPSTE24 vi LMNA. sin phim PCR sau iSl
Hinh 2. Dl$n d i s i n p h i m PCR nhSn 10 dogn exon c6a gen a»PSTE24. M: Mariier lOObp, 1-10. San phSm PCR fliSn di theo thip tir tip exon 1 d i n exon 10 cOa gen ZMPSTE24 K K v ",•
(A) AG A AC C GT ATC (B) AC C T NNK 7 ( C ) T T AT] TGtr GI ( D ) . ,
•j/\Jh.
Hlnh 3. So s d n h trinh nucleotide ciia hai dogn exon gen ZMPSTE24 vd trinh t y OA c6ng bO NM_005857. A. Trlnh t y dogn exon 1 cua gen ZMPSTE24 dgc t o sdn phdm PCR cda ngircri khoe mgnh. B. Trlnh t y dogn exon 1 cua gen ZMPSTE24 dgc tCr s i n p h i m PCR cua b$nh nhdn. C. Trlnh t y doan exon 9 cCia gen ZMPSTE24 age tir sdn p h i m PCR cCia ngiroi khoe mgnh D, Trlnh tir dogn exon 9 cua gen ZMPSTE24 dpc tCr sdn phdm PCR ciia b$nh nhan
S a u khi p h d n tfch v d s o s d n h trinh tif 12 d o g n e x o n c u a gen LMNA v&i t r i n h tif t r e n G e n B a n k ( N M _ 1 7 0 7 0 7 ) , c d c sdn p h i m P C R t h u d u ^ c dirg'c xdc (flnh c h f n h xac Id 12 d o g n e x o n cOa g e n LMNA, tuy nhidn k h d n g thu du-oc m$t thay d l i n u c l e o t i d e n d o .
Khi phdn tfch trinh ti,r g e n ZMPSTE24, k i t q u d thu d u ^ c 10 dogn e x o n c 6 c i i i ^ u d d i l l n l i j g i Id 2 6 8 bp, 399 bp, 232 bp, 2 7 3 b p , 2 3 1 b p , 300 bp. 292 b p , 2 2 9 bp, 2 8 4 bp, 3 1 9 b p t u ' o n g irng vi/i kfch t h i r d c tfnh todn ly t h u y l t ci^a exon 1 d i n e x o n 1 0 . Khi s o s d n h t r i n h tif c u a 10 dogn e x o n ndy viri t r i n h tu- g e n c i n g bo trdn G e n b a n k ( N M _ 0 0 5 8 5 7 ) , c h u n g t o i t h u d i r g e hai d | t b i l n d j h o p fa>, c.50de(A a e x o n 1 v d c . 1 0 8 5 - 1 0 8 6 i n s T it e x o n 9 ( H i n h 3) ie b d n h n h d n ky h i g u G - 1 0 6 8 8 . V I vdy, chOng toi t i l n hdnh d g c t r i n h t y tn,pc t i l p s a n p h i m PCR c u a hai e x o n 1 vd 9 cua c a b l m g b g n h n h d n . K i t q u a d p c trinh tif Inic t i l p ti> sdn p h i m P C R c h o t h i y b l (G-13725) v d m g ( G -
B-
2 G-mn
-o
cliaS-IMtinsT WT
c ^ d e U cIMS-IOItJDsT
Hinh 4. Pha h^ gia dinh Greifswald-RD'1.1.1: Bo b$nh nhSn (G- 13725) mang d$t b i l n dj hgp tir c.SOdelA. 1.2, M? b^nh nhan (G- ii'sic: k a „ ' k „ k A „ i A „ i,„™t ™ ^ „ „ " , * A t K : A / J k " ' . - 13625) mang dOt b i l n dj hop ti> c.1085-10e6insT. II, B^nh nh§n (G- 13625) b $ n h n h d n lan l u ' g l m a n g dOt b i l n d | h ^ p tu- loesS) mang cd hai dat b i i n di hgp to c.50 delA va c i o e s - c S O d e l A v d c . 1 0 8 5 - 1 0 8 6 i n s T ( H i n h 4 ) . 1066insT
Ndm 2 0 0 4 , d | t b i l n c . 1 0 8 5 - 1 0 8 6 i n s T d i u tidn d u g c t i m thay v a da d u p c c h t r n g m i n h la nguydn n h d n gdy b g n h c d n g d a ( N a v a n o ef al.. 2 0 0 4 ) . N 6 du-gc t i m t h i y b ^ n n h i l u b e n h n h d n c d n g da khdc (Navan-o ef al., 2 0 0 5 ) . D p t b i l n nay lam thay d l l k h u n g djch m a c u a protein tgo ra m $ t c h u l i peptWe n g l n h o n p.L362feX18. D o t b i l n cSOdelA cOng da d u v c c h i r n g m i n h Id n g u y e n n h d n gdy b g n h c d n g d a (Smigiel etaL, 2 0 1 0 ) . T u a n g t u n h u ' c . 1 0 8 5 - 1 0 8 6 i n s T , dot b i l n ndy c u n g lam thay d l i k h u n g d[ch m a tgo ra m $ t c h u o i peptide n g a n h o n p . K 1 7 ^ X 2 0 . D g t b i l n c.50delA da d u g c k i l m faB tren 3 8 6 allele c u a q u i n t h e n g i f o i b i n h t h u o n g d l iogi b o k h d n a n g ddy Id m g t da hinh nucleotide d o n . VI v d y c o c h l p h d n b>
gdy b § n h c d n g d a it b e n h n h d n G-10688 c 6 t h i du'gc giai t h i c h Id d o hai dgt b i l n c.50de[A v d c.1085-10B6insT o b B n hai allele k h d c n h a u c u a g e n ZMPSTE24 g d y n d n . Ca hai d i t b i l n ndy deu t g o ra c h u o i peptide ngdn h a n chuoi peptide binh t h u a n g {ddi 4 7 5 a m i n o acid) do d 6 Idm m a t c h u c n a n g cua pratein Z M P S T E 2 4 Khi k h o n g c6 mgt ciia e n z y m e Z M P S T E 2 4 , q u d t r i n h xi> ly s a u phidn m a cila prelamin A se k h i n g d i i n ra d l tao t h a n h lamin A c h i i c ndng t h a m gia v d o mOt s 6 hogt dOng t h i l t y e u ciia te bdo. T i l n lamin A se du-gc t i c h tg trang t l bao n h i l u h o n mil-c binh thu-ong. T i l n i a m i n A d a d u g c chCfng m i n h la c 6 khd n d n g gay d f l c va lam chet t l bdo (Navarro et al., 2 0 0 5 )
K C T L U A N
B d n g ky t h u d t P C R , d i u n g toi da nhan d u g c todn bp c a c d o a n exon cua c a hai g e n ZMPSTE24 vd LMNA cua b e n h n h d n c d n g da C d c d o g n e x o n ndy dd d u g c xdc dinh trinh t g t n / c t i l p . Qua p h a n t i c h k i t q u a d p c trinh t i / da thu d u w c hai dot b i e n dj h g p tu- t r o n g g e n ZMPSTE24. c 50delA di t m y l n t i r b o vd c 1085 - 1 0 8 6 i n s T di t r u y i n t u m e khi s o s d n h
•p?ill U 5 & ' - — ' - - , 6 i M n h t v g e n c o n g b6 tren G e n b a n k (NM_005857). (3 gen UANA kh6ng tlm t h i y dot b l i n nSo.
Lfricim tni
r.*__-j_tL • ,t L .^^.^MA,^. ,M^^hA,yrJiAr "Nphiin cuu ditrm^ phdn li cdc b4nhteo CIT &nery^Dre!fiiss(EDAm^
TAI LI^U THAM KHAO
Agarwal AK. Fryns JP. Audius RJ, Garg A (2003). Zinc metalioproteinase. ZMPSTE24, is mutated in mandibuloacral dysplasia. Hm Mol Genet 12(16): 1995-2001.
Ben Yaou R. Navano C. Quganofloy S. Bertrand AT, Massart C. De Sandre-Grovannoli A, Cadinanos J^ Mamchaoui K, Butler-Browne G. Estoumet B. Richard P, Baroe A. Levy N, Bonne G (2011). Type B mandibuloacral dysplasia with congenital myopathy due b homozygous ZMPSTE24 missense mutatk>n. Eur J Hum Genet 19(6): 647-654.
Eriksson M, Brown WT, Gordon LB. Glynn MW. Singer J. Scott L. Errios MR. Robbins CM. Moses TY. Berglund P, Dutra A, Pak E, Durkin S. Csoka AB, BoehiAe M, Gtover TW, Collins FS (2003), Recunent de novo point mutations in lamin A cause Hutchinson-Gilfoiti progena syndrome. Nature 423(6937): 293-298.
Holtz D. Tanaka RA, Hartwig J, McKeon F (1989). The CaaX motif of lamin A ftjnctions in conjunct'on wilh ttie nuclear localization signal to target assembly lo the nudear envetope Cell 59(6)' 969-977.
Ll C (2010) Homozygosity for Ibe common mutallon c.1085dupT in the ZMPSTE24 gene in a Mennonite baby with restnctive demiopathy and placenta abniptkxi Am J Med Genet A 152A(1): 262-263.
Moulson CL, Go G. Gardner JM, van der Wal AC. Smitt JH. -van Hagen JM, Miner JH (2005). Homozygous and compound heterozygous mutatons in ZMPSTE24 cause the laminopathy restrictive demiopathy. J Invest Demiatol 125(5). 913-919, Navarro CL. Cadinanos J, De Sandre^iovannoli A, Bernard R, Coumer S, Boccaccio I, Boyer A. Kleijer WJ, Wagner A, Giuliano F, Bewner F A Frelje JM, Cau P. Hennrfcam RC, Lopez-Otin C, Badens C. Levy N (2005). Loss of ZMPSTE24 (FACE-1) causes autosomal recessive restncbve demiopattiy and accumulation of Lamin A precursors Hum Md Genet 14(11): 1503-1513, Navarro CL, De Sandre-Gtovannoli A, Bernard R, Boccaccio I. Boyer A,-Genevieve D, Hadj-Rabia S, Gaudy-Marqueste C, Sm'itt HS, Vabres P. Faivre L, Verloes A Van Essen T, Flori E, Hennekam R, Beemer FA, Laurent N. Le Mener M. Cau P, Levy N (2004), Lamin A and ZMPSTE24 (FACE-1) detects cause nudear disorganization and identify restrictive demiopathy as a lethal neonatal laminopathy.
Hum Mol Genel 13(20), 2493-2503.
Prokocimer M. Davidovich M. Nissim-Ratinia M, Wlesei-Motiuk N, Bar DZ, Barkan R, Meshorer E, Groenbaum Y (2009), Nuclear lamins:
key regulators of nuclear structure and activities. J Cell Mol Med 13(6): 1059-1085.
Sinensky M, Fantle K, Trojillo M. McLain T, Kupfer A Dalton M (1994). The processing pathway of prelamin A. J Cell Sci 107 ( R 1)61- 67.
Smigiel R. J^utriak A Esteves-Vieira V, Szela K. Halon A, Jurek T, Levy N, De Sandre-Giovannoli A (2010). Novel frameshifling mutations of the ZMPSTE24 gene in two siblings affected wth restnctive dermopathy and review of the mutations described in tfie literature.AmJMedSenetA152A(2) 447-452.
SCREENING OF MUTATIONS RELATED TO RESTRICTIVE DERMOPATHY
N g u y e n T h u y D u o n g \ M a n f r e d W e h n e r t ^ , N o n g V a n Hai^
%stibJte of Human Research. Vietnam Academy of Science and Technology Ins&ute of Human Genetics. Greifswald, Germany
Restrictive dermopathy (RD) is a rare autosotnal recessive disorder caused by mutations in LMNA and ZMPSTE24 It is characterized by light and ngid skin with erosions, prominent superficial vasculature, RD afifected patienis noimaHy die soon after birth In lliis work, we screened mutations m two genes ZMPSTE24 and LMNA for a family affected with RD. Using PCR and specific pnmm.
we codd amplify «" "^JfJ-f »hc two genes TTien, PCR products were punfied and sequenced. Sequence alignment results showed J w p ^ C ^ i T f ^ a T ^ '«="°^e««e of cSOdelA and c,I085-1086insT in ZMPSTE24 when compared witli
^ ^ 1 \ M 1 £ ^ ' ^ ^ ^ >«4aceessioi. nun,b^NM^005857. Tbe heterozygous c.SOdelA inherited from the father was not m ^ S ta ! l S H ^ r ^ ^ " ^ ^°^ T ? ' "^^^^eous nmtalion c.1085 -1086msT was common and found in the Z ^ T O T ^ e i w T L ^ I ™ " " " ^ ^ " - f ^"^^^ =°-P^«d with LMNA published in GenBank with accession number NM_170707. TTie result will parUy conmbute to molecular studies on human inherited disorders.
Key words: LMNA, mutation, PCR, Resuictive dermopathy, ZMPSTE24
