WO1998031835A1 - Genetic methods for identifying individuals for improving well being and performance through exercise - Google Patents

Abstract

Methods for improving the health and/or performance of a person having identified genotypes through exercise.

Description

GENETIC METHODS FOR IDENTIFYING INDIVIDUALS FOR IMPROVING WELL BEING AND PERFORMANCE THROUGH EXERCISE

GROSS-REFERENCE TO RELATED APPLICATIONS

This application is a non-provisional application claiming benefit of provisional application No. 60/035,382, filed January 16, 1997 and 60/048,309, filed May 27, 1997, both applications herein incorporated by reference.

FIELD OF THE INVENTION

This invention relates to identifying alleles associated with phenotypes and recommending lifestyle changes based on the allele identification. BACKGROUND OF THE INVENTION

Studies have shown that individuals suffering from hypertension, insulin resistance, arthritis, unfavorable cholesterol profiles, and other ailments can alleviate their symptoms or improve their conditions through exercise. Unfortunately, some individuals, no matter how rigorously they exercise or train, are unable to improve their conditions, and yet still others benefit to a much greater extent than predicted.

These results underscore the fact that many factors contribute to one's well-being. Such factors include, for instance, behaviors such as diet and exercise, genetic makeup, and environment .

While behavior can be controlled or altered, and our environments can be regulated, our genetic makeup, at least up until now, is predetermined and set at birth. By identifying the genetic makeup of a population and recognizing that some individuals of the population will benefit from a change of behavior, and still others will benefit to a much greater extent, and still others do not benefit at all, one hypothesizes that the genotype of an individual is a predictor of a result achieved through a change of behavior.

Thus, an object of this invention is to identify individuals possessing a certain genotype and associated ailment, and to determine if the health of that individual can be improved by altering behavior.

Another object of the invention relates to identifying individuals having certain apolipoprotein (APO) E and lipoprotein (LPL) genotypes and recommending present or future changes in behavior that will improve the cholesterol profile of such person.

Another object of the invention relates to identifying patients having certain angiotensin converting enzyme (ACE) , APO E and LPL alleles and recommending present or future behavioral changes that will improve hypertension in that individual .

Another object of the invention relates to identifying women having certain Vitamin D receptor (VDR) alleles and recommending present or future changes in behavior that will improve the bone mineral density of such women. Another object of the invention relates to identifying persons having certain ACE alleles for the purpose of quantifying their current and future risk of developing cardiovascular disease. DETAILED DESCRIPTION OF THE INVENTION For purposes of this invention a lifestyle change may involve a course of action to overcome or enhance a particular genotype. The course of action may be, for example, a change in diet, starting an exercise program or both, a change in environment, or for instance, changing an exercise regime, etc . Cholesterol and APO E

Apolipoprotein E (APO E) [SEQ. ID NO.2] plays a central role in cholesterol transport, total cholesterol and low- density lipoprotein cholesterol concentrations, and contributes to coronary heart disease risk. APO E is a ligand for lipoprotein receptors. Physiologically, its most important function is to mediate specific uptake of plasma very low-density lipoproteins, chylomicron remnants, and intermediate-density lipoprotein by the liver. The APO E gene disclosed in Hixson et al, "Restriction isotyping of human apolipoprotein E by gene amplification and cleavage with Hhal." Journal of Lipid Research Volume 31, 1990 (herein incorporated by reference) [SEQ. ID NO. 1], is polymorphic with three common alleles designated E2 , E3 , and E4 , resulting in six major Apo E genotypes: E2/2, E3/2, E4/2, E3/3, E4/3 and E4/4. The Apo E3 allele is the most common allele in populations studied. Isoform E3 is distinguished by cysteine at position 112 (112 cys) and arginine at codon 158 in the receptor binding region of Epo E. Codon 158 is a positively charged region of the molecule that binds to low-density lipoprotein receptors. The E4 isoform (112arg and 158arg) is associated with increased levels of total cholesterol and betalipoprotein. The E4 allele has an amino acid substitution at Codon 112 that has arginine in place of cysteine, and this region appears to reduce disulfide bonding of Apo E with other free sulfhydryl-containing proteins. Consequently, APO E4 may be more readily transferred from high-density lipoprotein to chylomicron remnants than APO E3 resulting in enhanced receptor-mediated clearance of low-density lipoprotein, hepatic cholesterol accumulation, down-regulation of lipoprotein receptors, and subsequently raised concentration of serum cholesterol, giving rise to increased susceptibility to heart disease. Most patients with type III hyperlipidemia (a condition characterized by the accumulation in plasma of remnants of the metabolism of triacylglycerol-rich lipoproteins) , are homozygous for the E2 isoform (112cys and 158cys) that binds with reduced affinity to cellular receptors. In population studies, the E2 isoform is associated with decreased levels of cholesterol and betalipoprotein .

Lipoproteins with Apo E2 bind less well to low-density lipoprotein receptors, so clearance is slower. Low-density lipoprotein receptors are subsequently up-regulated, and serum cholesterol concentrations are reduced in the majority of individuals with the E2 allele. However, 1-10 % of individuals with the E2/2 genotype develop type III hyperlipidemia. This variability in response suggests that other genes and/or environmental factors play important roles in the development of overt type III hyperlipidemia in subjects with the E2/2 genotype.

The APO E polymorphism has been consistently shown to be associated with plasma concentrations of total and low-density lipoprotein cholesterol.

Identifying Individuals with Apo E Alleles. In a first study, obese sedentary hypertensive men, 50-65 years of age, had DNA collected from them, and this DNA was analyzed for the presence of the Apo E alleles identified by the method below:

Typing of the subjects was determined by using an isoelectric focusing-immunoblotting method as described previously Kamboh et al, "Impact of apolipoprotein E polymorphism in determining interindividual variation in total cholesterol and low density lipoprotein cholesterol in Hispanics and non-Hispanics whites". Atherosclerosis 1993;98:201-11 (herein incorporated by reference). Other methods for identifying APO E alleles include using the polymerase chain reaction (PCR) see Kamboh et al, "The relationship of APO E polymorphism and cholesterol levels in normoglycemic and diabetic subjects in a biethnic population from the San Luis Valley, Colorado." Atherosclerosis 1195:112:145-149. DNA was extracted from lymphocytes as described by Miller et al, "A simple salting out procedure for extracting DNA from human nucleated cells." Nucleic Acids Research 1988:16:1215. Genomic DNAs (0.5-1.0 μg) are amplified by using a forward primer El, 5'-GCGGACATGGAGGACGTG- 3' [SEQ. ID NO. 3] (codons 106-111) and a reverse primer E2 , 5'- GGCCTGGTACACTGCCAG-3' [SEQ. ID NO. 4] (codons 159-164) . The 50-μL reaction mixture consisted of 5 μL 10 x reaction buffer (100 mmol Tris-HCl/L, pH 8.9, 500 mmol KCl/L, 15mmol MgCl₂/L, 3 μL dimethylsulfoxide, 0.75 μL (0.3 μmol) of each primer; 0.6 μL each of (lOmmol/L) dATP, dGTP, dCTP, and dTTP; 0.3 μL Thermus aqua ticus ( Taq) DNA polymerase (Perkin Elmer Cetus

Inc., Foster City, CA) ; and 37.8 μL sterile deionized water, which was added to the DNA template. Twenty- five microliters mineral oil was placed over the final PCR reaction mixture. After a denaturation step of 8 min at 95°C, amplification is achieved by 30 cycles of denaturation (1 min at 95°C) , annealing (1 min at 57°C) , and extension (2 min at 72°C) , followed by extension for 5 min at 72°C. The nucleotide amplified product is digested directly with the restriction enzyme H al (New England Bio Labs, Beverly, MA) . The digested

DNAs are separated on 8% nondenaturing polyacrylamide gel in 1 x Tris borate EDTA buffer followed by staining with ethidium- bromide solution and the APO E polymorphism was typed by visualization under ultraviolet light.

These subjects were exposed with aerobic exercise training and were given the same diet. Specifically, to eliminate the effect of diet, all subjects were instructed in the principles of weight-maintaining American Heart Association (AHA) step I diet over an 8 -week period before baseline testing. This diet consisted of 50-55% of calories as carbohydrate, 30-35 % as fat, 15-20% as protein, 300-350 mg/day of cholesterol, and 3 g/day of sodium. These subjects were counseled weekly to maintain their diet consumption throughout the length of monitoring. Adherence was monitored by registered dieticians who reviewed weekly food records and body weights and calculated dietary consumption from biweekly 7-day food records. At baseline and after the intervention, the subjects were weight-stable for 4 weeks before testing. During this period, the subjects were instructed to maintain their body weight within 1 kilogram.

These subjects took part in an aerobic exercise program that met 3 times per week. Exercise training consisted of stationary cycling and walking and jogging on a treadmill starting at 50-60% of each individual's heart rate reserve for three five- to ten-minute periods. Target heart rate was calculated for each individual with the equation of Karvonen et al . , "The effects of training heart rate: a longitudinal study". Ann. Med. Exp . Biol . Fenn . 35:307-315 (1957).

Training intensity was gradually increased by five to ten percent of the heart rate reserve every month. At three months, the maximal oxygen consumption (V0_2max) test was repeated, and the intensity was adjusted until forty minutes of training per session at an intensity of seventy-five to eighty-five percent of heart rate reserve was achieved. All training sessions were supervised by the research staff, and the subjects were instructed by a dietician to increase the caloric intake to offset the increase in energy expenditure due to the increased physical activity.

Serum total cholesterol and high-density lipoprotein cholesterol and high density lipoprotein 2 cholesterol levels were measured for the subjects. Subjects possessing at least one APO E2 allele increased their high-density lipoprotein cholesterol (HDL-C) levels over seven times more with exercise training than those with only APO E3 or E4 alleles (Table 1) .

High-density lipoprotein cholesterol 2 (HDL₂-C) increases with exercise training were also dramatically larger in APO E2 vs.

APO E3 and E4 individuals (Table 1) .

Table 1: Plasma Lipoprotein-lipid changes with exercise training as a function of genotype TABLE 1

"n"= the number of subjects in each group

"p" indicates statistical probability for the difference between group

In a second study, subjects with lipoprotein lipase PvuII- /- genotype also had substantially greater exercise training- induced increases in plasma HDLC and HDL₂C levels than those with +/+, or +/- genotypes (Table 2) .

Lipoprotein lipase (LPL) [SEQ. ID NO. 6] is an enzyme that catalyzes the breakdown of triglycerides in the plasma to release free fatty acids. This hydrolysis also influences the metabolism of circulating lipoproteins. LPL has also been shown to enhance the triglyceride-rich chylomicron binding to low density lipoprotein receptor-related proteins. Thus, LPL may also be an important regulator of chylomicron metabolism. The LPL gene [SEQ. ID NO. 5] is located on human chromosome 8p22. It is approximately 35 kilobases long and has 10 functionally differentiated exons . Two primary polymorphic variations occur at the LPL gene locus in frequencies that are important on a population basis; these two markers are detected by PvuII and HinduI. There are three genotypes at each site with the alleles for both denoted as "+" or "-" based on the presence or absence of a restriction site at the LPL locus with PvuII or Hindlll. Thus, for both PvuII and HindiII there are three genotypes : +/+, +/-, and -/-. PvuII polymorphic variations have been reported to be associated with variations in plasma triglyceride levels. Hindlll variations have previously been shown to be associated with hypertriglyceridema, coronary artery disease, plasma total and HDL-cholesterol levels, and plasma insulin levels.

One individual with the APO E2 and LPL PvuII -/- genotypes elicited the largest HDL-C and HDL₂-C increases of the nineteen men in the study. Training-induced changes in total cholesterol and triglyceride levels were similar in all APO E and LPL PvuII genotype groups. No other training-induced changes in variables that affect plasma lipoprotein-lipids (including dietary habits, ^v°2_maχ body weight, and body composition) differed among either APO E or LPL PvuII genotype groups. Thus, these results show that APO E or LPL PvuII genotype identifies individuals who will improve their blood cholesterol profile with exercise training. TABLE 2

"n"= the number of subjects in each group

"p" indicates statistical probability for the difference between group

In a third study, cross-sectional differences in plasma HDL- C and HDL₂-C levels were found that provide further evidence that gene markers identify individuals most likely to improve their blood cholesterol profile with exercise training (Table 3) . Postmenopausal women were studied to assess the effect of physical activity behavior on cardiovascular disease risk factors .

Women were classified as postmenopausal by self-reported lack of menses for greater than two years and elevated levels of follicle stimulating hormone and luteinizing hormone. Women were classified as sedentary if they had not participated in regular aerobic activity for greater than two years. Women who participated in aerobic exercise for greater than 90 minutes/week, for greater than three years, but who were not training for endurance-based competitive events were classified as physically active.

Endurance trained women were defined as those undergoing rigorous exercise training more than 4-5 times per week for at least two years in preparation for competitive endurance-based events (primarily long distance running) .

It was found that the APO E genotype did not affect plasma lipoprotein lipids in either sedentary or physically active women. However, among endurance-trained women, those with at least one APO E2 allele had substantially higher HDL-C and HDL₂~C levels than those with only APO E3 or E4 alleles. Furthermore, endurance-trained women with the APO E2 genotype had higher HDL-C and HDL₂-C levels than the sedentary or physically active women, whereas HDL-C and HDL₂-C levels were the same in all APO E3 or E4 women regardless of physical activity habits. No other differences were evident between the APO E genotype groups of endurance-trained women that could affect plasma lipoprotein lipids including dietary habits, V0_2max, body weight, body composition, regional distribution of fat, running mileage, years of running, years postmenopausal, and hormone replacement status. Thus, these data also show that genetic markers identify individuals most likely to improve their blood cholesterol levels with exercise training.

Table 3: HDL-C and HDL₂-C levels as a function of genotype and exercise training status

The values reported are mean values for each group n= the number of subjects in a group

The difference between the two genotype groups within the endurance-trained women is p=0.20 for HDL-C and p=0.035 for HDL₂-C. The difference between endurance-trained women and the combined group of sedentary and physically-active women within the APO E 2/2 and 2/3 genotype is p=0.15 for HDL-C and p=0.08 for HDL₂-C.

Thus, high levels of physical activity interact with a woman's genotype such that only well-trained postmenopausal women with the APO E 2/2 or 2/3 genotype have better HDL-C and HDL₂-C levels compared to their sedentary or physically active peers .

With this knowledge, clinical diagnostic kits made available can be used in identifying individuals most likely to improve their blood cholesterol profiles with exercise training. Hypertension and ACE ACE

Angiotensin converting enzyme (ACE) [SEQ. ID NO . 8] is the enzyme responsible for catalyzing the conversion of angiotensin I, a relatively inactive tissue and plasma vasopressor hormone, into the potent and highly active vasopressor hormone, angiotensin II. This cascade of reactions is part of the renin- angiotensin-aldosterone system that has long been known to be an important regulator of arteriolar relaxation and vasoconstriction, and hence blood pressure, in humans and animals. The ACE gene [SEQ. ID NO. 7] is polymorphic with two common alleles designated I and D, resulting in three genotypes: II, ID, and DD . The D allele has a 287-base pair marker in intron 16 of the ACE gene deleted, whereas the I allele has the 287-base pair marker inserted. The D allele is associated with increased levels of ACE in both plasma and ventricular tissues. Increased levels of ACE will clearly contribute to increased myocardial and vascular smooth muscle growth and increased arteriolar vasoconstriction. Thus, the presence of the D allele would by hypothesized to have deleterious effects on the cardiovascular system and, in fact, the D allele has been associated with increased risk of left ventricular hypertrophy, cardiovascular disease, and sudden cardiovascular death. The D allele was also originally believed to be more prevalent in hypertensives than normotensives ; however, this relationship has generally not been substantiated in more recent studies. Identifying Individuals with Different ACE Alleles to Predict Blood Pressure Changes with exercise Training Obese sedentary hypertension men, 50-65 years of age had DNA collected from them and the DNA was analyzed for the presences of ACE alleles. Typing of these individuals was conducted by isolating high molecular weight genomic DNA from whole blood mononuclear cells by the procedure of Miller et al (1988) . ACE genotyping was carried out by polymerase chain reaction amplification using the forward primer 5'- CCGTTTGTGCAGGGCCTGGCTCTCT-3 ' [SEQ. ID No . 9 ] and reverse primer 5' -CAGGGTGCTGTCCACACTGGACCCC-3 ' [SEQ. ID NO.10 ] and the following cycling conditions: denaturation at 95°C for 5 minutes followed by thirty cycles of 30 sec. Denaturation at 94°C, 15 sec annealing at 58°C, 30 sec. extension at 72°C. Amplimers were resolved on 2% agarose gels and genotypes assigned by direct comparison to samples of known genotype. The I (insertion) allele yielded a band of 490 bp and the D (deletion) allele a band of 190 bp. Heterozygotes were typed by the presence of both bands plus a heteroduplex band migrating at an approximately 370 bp . (Tiret et al . "Evidence from combined segregation and linkage analysis that a variant of the angiotensin-1 converting enzyme (ACE) gene controls plasma ACE levels" AM J. Hum Genet 1992; 51:197-205) reported above.

These subjects were exposed with aerobic exercise training and were given the same diet. All subjects were instructed in the principles of weight-maintaining American Heart Association step I diet over an 8 -week period before baseline testing as reported at page 6. These subjects were counseled weekly to maintain their diet as reported at page 6. Adherence was monitored as reported at page 6.

These subjects took part in an aerobic exercise program that met 3 times per week as reported at pages 7.

Subjects had their blood pressure measured weekly for 4 weeks prior to and following the completion of the exercise training intervention. Blood pressure measurements were made using a mercury sphygmomanometer and stethoscope according to standards established by the American Heart Association for cuff size, Korotkoff sounds, pre-measurement rest, and the number, timing, and averaging of the blood pressure results. The final values at baseline and after the intervention represent the average of 12 independent blood pressure measurements (3 on each of the 4 measurement days) .

Results of this study indicate that ACE genotype identifies hypertensive individuals that reduce their systolic and diastolic blood pressure with exercise training. Those subjects with at least one insertion ("I") ACE allele decreased their diastolic blood pressure with exercise training approximately 7 times more than those with only deletion ("D") ACE alleles (Table 4) . Subjects with at least one insertion ACE ("I") allele decreased their systolic blood pressure with exercise training over twice as much as those with only deletion ACE ("D") alleles (Table 4) . Subjects' baseline characteristics prior to exercise training did not differ among ACE genotype groups. Furthermore, no other training-induced changes in variables that affect systolic and diastolic blood pressure, including dietary habits, V0₂max, body weight, and body composition, differed among ACE genotype groups.

Thus, these results show that ACE genotype is a strong independent indicator of those individuals who will reduce their systolic and diastolic blood pressure with exercise training.

Table 4 : Change in Blood Pressure with Exercise Training in Hypertensives as a Function of ACE Genotype

In yet another method, applicants have discovered that individuals with different LPL PvuII genotypes, after exercise training will exhibit different results relative to systolic and diastolic blood pressure not predicted from initial screenings.

Specifically, the same obese sedentary hypertensive men age

50-65 earlier described had DNA collected from them, and this DNA was analyzed for the presence of genetic variations at two critical restriction sites at the lipoprotein lipase (LPL) gene locus. (See page 9 supra)

DNA samples were subjected to amplification by the polymerase chain reaction in a Perkin-Elmer Cetus DNA Themal

Cycler. One set of primers was derived from sequences between exons 8 and 9 in the, LPL gene to amplify the sequence around a

Hindlll restriction site in intron 8 (the forward primer was 5'- TTTA GGCCTGAAGTTTCCAC-3 ' [SEQ ID NO. 11] and the reverse primer was 5'CTCCCTAGAAGAGAAGATC-3 ' [SEQ ID NO. 12] as described

(Kirchgessner TG. et al . , "Organization of human lipoprotein lipase gene and evolution of the lipase gene family." Proc National Academy of Science 86: 9647-9651, 1989). The amplified fragment had a size of 1.3 kilobases. The second set of primers was from the DNA sequences flanking the PvuII reaction site. In intron 6 (the forward primer was 5 ' TAGGAGGTTGAGGCACCTGTGC-3 '

[SEQ. ID NO. 13] and the reverse primer was 5'GTGGGTGAATCACCTGAGGTC-3 ' [SEQ. ID NO. 14 ] as described (Oka

K et al . , "Nucleotide sequence of PvuII polymorphic site at the human lipoprotein lipase gene locus." Nucleic Acid Research 17:

6752, 1989) . This amplified fragment was 858 base pairs long.

The 50 ul reaction mixture contained 1 x PCR buffer (lOmM Tris, pH 8.3. 50mM KCl, 1.5mM MgCl₂,), dNTPs at 200uM, 0.3uM each primer, 0.5 ug genomic DNA, and 1.25 units of Taq DNA polymerase. Amplification of the region flanking the Hindu site was carried out for 33 cycles at 95°C for 1 min, at 60°C for 2 min, and at 72° for 2 min. For amplification around the PvuII site, the conditions were the same except for annealing at 70°C and 25 cycles. Amplified products were digested with Hindlll or PvuII and the resulting fragments separated on 2% agarose gels. After digestion with, Hindlll the presence of the restriction site (+ allele) resulted In fragments of 600 and 700 base pairs. The Presence of the PvuII site (+ allele) yielded fragments of 266 and 592 base pairs.

These subjects were exposed with aerobic exercise training and were given the same diet as reported at page 6. Specifically, to eliminate the effect of diet, all subjects were instructed in the principles of weight-maintaining American Heart Association step I diet over an 8 -week period before baseline testing as reported at page 6. These subjects were counseled weekly to maintain their diet consumption throughout the length of monitoring. Adherence was monitored as reported at page 10.

These subjects took part in an aerobic exercise program as reported at pages 7.

Prior to having undergone exercise training and after undergoing the exercise regimen and associated diet (the same regimen and diet as reported at page 6 above) , subjects were tested for systolic and diastolic blood pressure levels. Initial levels did not differ between hypertensive individuals with different LPL PvuII genotypes. However, the changes in blood pressure resulting from exercise training did differ among LPL PvuII genotype groups (see Table 5) . Those having only + alleles decreased both their systolic and diastolic blood pressures more with exercise training than those with the -/- or the +/- genotype. In fact, both genotype groups decreased their systolic and diastolic blood pressure significantly with exercise training. However, those men with the +/+ LPL PvuII genotype tended to decrease both their systolic and diastolic blood pressure more than men with the +/- or -/- genotype. The subjects baseline characteristics prior to exercise training did not differ among LPL PvuII genotype groups, except that the -/- and the +/- genotype men were somewhat older that the +/+ men (63.8 vs 56.5 years). Furthermore, no other training-induced changes in variables that affect systolic and diastolic blood pressure, including dietary habits, V0₂max, body weight, and body composition, differed among LPL PvuII genotype groups.

TABLE 5

Changes in blood pressure with exercise training in the two

LPL PvuII genotype groups

"n"= the number of subjects in each group

Thus, these results indicate that LPL PvuII genotype is an indicator of those individuals most likely to reduce their systolic and diastolic blood pressure the most with exercise training.

In these same individuals, initial systolic blood pressure levels did not differ between hypertensive individuals with different LPL Hindlll genotypes, but initial diastolic blood pressures were somewhat higher in LPL Hindlll +/+ and +/- genotype compared to -/- genotype men (94 vs. 86 mmHg) . However, the changes in blood pressure resulting from exercise training did differ among LPL Hindlll genotype groups (see Table 6.) Those having at least one + allele decreased both their systolic and diastolic blood pressures significantly with exercise training. Furthermore, men with the +/+ or +/- LPL Hindlll genotype decreased both their systolic and diastolic blood pressures more than men with the -/- genotype. The subjects' other baseline characteristics prior to exercise training did not differ among LPL Hindlll genotype groups. Furthermore, no other training-induced changes in variables that affect systolic and diastolic blood pressure, including dietary habits, V0₂max, body weight, and body composition, differed among LPL Hindlll genotype groups .

TABLE 6 Changes in blood pressure with exercise training in the two LPL Hind III genotype groups

"n"= the number of subjects in each group

Thus, these results indicate that LPL Hindlll genotype is an indicator of those individuals most likely to reduce their systolic and diastolic blood pressure the most with exercise training.

Vitamin D receptor gene [SEP. ID NO. 151 and Bone density The vitamin D receptor [SEQ. ID NO. 16] plays a central role in the regulation of calcium metabolism, hence it also has a critical role in determining bone homeostasis. The Vitamin D receptor combines with other nuclear factors to activate the transcription of a large number of target genes including osteocalcin. The vitamin D receptor has been found to have a polymorphic variation that is detected with BsmI . Alleles with the presence of the BsmI restriction site are denoted as "b" and those without the BsmI restriction site are denoted as "B" giving rise to three distinct vitamin D receptor genotypes: bb, Bb, and BB. These genotypes have previously been found to be related to circulating osteocalcin levels and bone mineral density in both young and postmenopausal women. Furthermore, these genotypes have been found to be related to risk of hip fracture and the rate of bone loss after the menopause in women.

In order to identify these alleles, high molecular weight DNA was extracted from peripheral leukocytes by standard methods. (SA Miller et al . A simple salting out procedure for extracting, DNA from human nucleated cells. Nucleic Acids Research 16: 1215-1218, 1988). DNA was amplified by PCR, using amplification primers as described by NA Morrison, et al . "Contribution of trans-acting factor alleles to normal physiologic variability, vitamin D receptor gene polymorphism and circulating osteocalcin." Proc National Acad Sci 89: 6665-6669, 1992) . Each PCR was performed using 60 ul final reaction volume containing 110-200 ng DNA, 0.46 uM of each primer, 185 uM of dNTP mixture, 50mM KCI, 10 mM Tris-HCI pH 9.0, 1.5 mM MgCl, 0.1% Triton X-100, 0.8 units Taq polymerase). After amplification, 5 units of BsmI (New England Biolabs, Beverly, MA) was added to 16 uL of amplified product for digestion at 65° C. Each digested sample was loaded onto 2% agarose gels containing ethidium bromide and electrophoresed for 3 hours at 90 volts, After electrophoresis , the DNA fragments were visualized by UV illumination and fragment sizes were estimated by comparison to a 1 kilobase size ladder run on the same gel. The presence of a polymorphic restriction site was specified as "b", whereas the absence of this site was specified as "B" .

Fifty-one healthy postmenopausal women were recruited into six specific groups based on their hormone replacement therapy status and habitual levels of activity. Women were classified as postmenopausal by self-reported lack of menses for greater than two years and elevated levels of follicle stimulating hormone and luteinizing hormone. Women were classified as sedentary if they had not participated in regular aerobic activity for greater than two years. Women who participated in aerobic exercise for greater than 90 minutes/week, for greater that three years, but who were not training for endurance-based competitive events were classified as physically active. Endurance trained women were defined as those undergoing rigorous exercise training more than 4-5 times per week for at least two years in preparation for competitive endurance-based events (primarily long distance running) .

Hip and lumbar spine bone mineral density were determined on all women in the morning after an overnight fast while wearing standard hospital gowns using a Lunar Corporation DPX-L dual energy X-ray absorptiometry system. Scans were obtained and analyzed using conventional methods and software. Quality control measures were recorded prior to each scan to ensure validity of the bone mineral density result.

The results of testing for the bone density of the women are reported below: TABLE 7 TROCHANTER BONE MINERAL DENSITY

Values are means expressed in g/cm²

Hormone replacement therapy had no effect on bone mineral density in these women. These results show that low to moderate levels of habitual physical activity interact with the BB vitamin D receptor resulting in increased bone mineral density of postmenopausal women.

The ACE locus and VQ₂max

In yet another study healthy postmenopausal women 50-75 years of age underwent V0₂max testing on a treadmill to determine the maximal amount of oxygen they could consume (V0₂max) when exercising to their own individual maximal capacity. The women were almost equally divided between women on and not on hormone replacement therapy. In addition, the women were approximately equally divided between groups that were sedentary, physically- active, and endurance trained. Hormone replacement therapy did not affect V0₂max. Habitual physical activity level affected V0₂max with the sedentary women having the lowest, the physically active having intermediate, and the endurance trained women having the highest V0₂max values. ACE genotype also affected V0₂max in these women after accounting for the effects of habitual physical activity level on V0₂max see Table 8 below: TABLE 8

V0₂max values in postmenopausal women with different habitual physical activity levels as a function of ACE genotype.

Values in Table 8 are expressed as mean ± standard deviation in units ml/kg/min. Values in parentheses indicate the number of subjects in each group. To account for the different physical activity levels of the women, which independently affects V0₂max, each woman's V0₂max was expressed as a difference from the average value for their respective habitual physical activity level group (sedentary, physically-active, endurance-trained) . Thus, positive values indicate results above the average based on physical activity level and negative values indicate results below the average based on physical activity level.

Both the II and DD genotype groups were significantly different from zero (P=0.04 and 0.02 respectively), whereas the ID genotype group was clearly not significantly different from zero. The difference between II and DD genotype groups was also statistically significant (P=0.02). The differences between the two homozygote groups when compared to the heterozygote ID genotype group both approached statistically significant (p=0.14- 0.15) .

These results show that a specific gene marker can identify an individual having a higher or lower V0₂max value compared to their peers. Thus, an individual with an ACE II genotype can be directed and would be expected to have the ability to compete with great success in activities such as running, cycling and swimming. Show and performance animals such as steeplechase horses and thoroughbreds, and greyhounds etc. having a similar genotype would also be expected to have the ability to compete with great success.

Because a low V0₂max is a risk factor for cardiovascular disease, ACE genotype will also help to identify individuals at lower and greater risk for cardiovascular disease.

SEQUENCE LISTING

(1) GENERAL INFORMATION:

(i) APPLICANT: HAGBERG, JAMES M FERRELL, ROBERT E

(ii) TITLE OF INVENTION: GENETIC METHODS FOR IDENTIFYING INDIVIDUALS FOR IMPROVING WELL BEING OR PERFORMANCE

THROUGH EXERCISE

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(C) STRANDEDNESS : single (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

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(xi) SEQUENCE DESCRIPTION: SEQ ID NO:l:

TAAGCTTG GCA CGG CTG TCC AAG GAG CTG CAG GCG GCG CAG GCC CGG CTG 50 Ala Arg Leu Ser Lys Glu Leu Gin Ala Ala Gin Ala Arg Leu 1 5 10

GGC GCG GAC ATG GAG GAC GTG CGC GGC CGC CTG GTG CAG TAC CGC GGC 98 Gly Ala Asp Met Glu Asp Val Arg Gly Arg Leu Val Gin Tyr Arg Gly 15 20 25 30

GAG GTG CAG GCC ATG CTC GGC CAG AGC ACC GAG GAG CTG CGG GTG CGC 146 Glu Val Gin Ala Met Leu Gly Gin Ser Thr Glu Glu Leu Arg Val Arg 35 40 45 CTC GCC TCC CAC CTG CGC AAG CTG CGT AAG CGG CTC CTC CGC GAT GCC 194 Leu Ala Ser His Leu Arg Lys Leu Arg Lys Arg Leu Leu Arg Asp Ala 50 55 60

GAT GAC CTG CAG AAG CGC CTG GCA GTG TAC CAG GCC GGG GCGAATTCTG 243 Asp Asp Leu Gin Lys Arg Leu Ala Val Tyr Gin Ala Gly 65 70 75

244

(2) INFORMATION FOR SEQ ID NO : 2 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 75 amino acids (B) TYPE: amino acid

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 2 :

Ala Arg Leu Ser Lys Glu Leu Gin Ala Ala Gin Ala Arg Leu Gly Ala 1 5 10 15 Asp Met Glu Asp Val Arg Gly Arg Leu Val Gin Tyr Arg Gly Glu Val

20 25 30

Gin Ala Met Leu Gly Gin Ser Thr Glu Glu Leu Arg Val Arg Leu Ala 35 40 45 Ser His Leu Arg Lys Leu Arg Lys Arg Leu Leu Arg Asp Ala Asp Asp 50 55 60

Leu Gin Lys Arg Leu Ala Val Tyr Gin Ala Gly 65 70 75

(2) INFORMATION FOR SEQ ID NO : 3 :

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS : single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO (iv) ANTI- SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: GCGGACATGG AGGACGTG 18

(2) INFORMATION FOR SEQ ID NO : 4 : (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 18 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:

GGCCTGGTAC ACTGCCAG 18

(2) INFORMATION FOR SEQ ID NO : 5 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3549 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic)

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Homo sapiens

(F) TISSUE TYPE: Adipose tissue

(ix) FEATURE:

(A) NAME/KEY: sig_peptide

(B) LOCATION: 175..255

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 175..1602 (ix) FEATURE:

(A) NAME/KEY: mat_peptide

(B) LOCATION: 256..1599

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5:

CCCCTCTTCC TCCTCCTCAA GGGAAAGCTG CCCACTTCTA GCTGCCCTGC CATCCCCTTT 60 AAAGGGCGAC TTGCTCAGCG CCAAACCGCG GCTCCAGCCC TCTCCAGCCT CCGGCTCAGC 120

CGGCTCATCA GTCGGTCCGC GCCTTGCAGC TCCTCCAGAG GGACGCGCCC CGAG ATG 177

Met -27 GAG AGC AAA GCC CTG CTC GTG CTG ACT CTG GCC GTG TGG CTC CAG AGT 225 Glu Ser Lys Ala Leu Leu Val Leu Thr Leu Ala Val Trp Leu Gin Ser -25 -20 -15

CTG ACC GCC TCC CGC GGA GGG GTG GCC GCC GCC GAC CAA AGA AGA GAT 273 Leu Thr Ala Ser Arg Gly Gly Val Ala Ala Ala Asp Gin Arg Arg Asp -10 -5 1 5

TTT ATC GAC ATC GAA AGT AAA TTT GCC CTA AGG ACC CCT GAA GAC ACA 321 Phe lie Asp lie Glu Ser Lys Phe Ala Leu Arg Thr Pro Glu Asp Thr 10 15 20

GCT GAG GAC ACT TGC CAC CTC ATT CCC GGA GTA GCA GAG TCC GTG GCT 369 Ala Glu Asp Thr Cys His Leu lie Pro Gly Val Ala Glu Ser Val Ala 25 30 35

ACC TGT CAT TTC AAT CAC AGC AGC AAA ACC TTC ATG GTG ATC CAT GGC 417 Thr Cys His Phe Asn His Ser Ser Lys Thr Phe Met Val lie His Gly 40 45 50 TGG ACG GTA ACA GGA ATG TAT GAG AGT TGG GTG CCA AAA CTT GTG GCC 465 Trp Thr Val Thr Gly Met Tyr Glu Ser Trp Val Pro Lys Leu Val Ala 55 60 65 70 GCC CTG TAC AAG AGA GAA CCA GAC TCC AAT GTC ATT GTG GTG GAC TGG 513 Ala Leu Tyr Lys Arg Glu Pro Asp Ser Asn Val lie Val Val Asp Trp 75 80 85

CTG TCA CGG GCT CAG GAG CAT TAC CCA GTG TCC GCG GGC TAC ACC AAA 561 Leu Ser Arg Ala Gin Glu His Tyr Pro Val Ser Ala Gly Tyr Thr Lys 90 95 100 CTG GTG GGA CAG GAT GTG GCC CGG TTT ATC AAC TGG ATG GAG GAG GAG 609 Leu Val Gly Gin Asp Val Ala Arg Phe lie Asn Trp Met Glu Glu Glu 105 110 115

TTT AAC TAC CCT CTG GAC AAT GTC CAT CTC TTG GGA TAC AGC CTT GGA 657 Phe Asn Tyr Pro Leu Asp Asn Val His Leu Leu Gly Tyr Ser Leu Gly 120 125 130

GCC CAT GCT GCT GGC ATT GCA GGA AGT CTG ACC AAT AAG AAA GTC AAC 705 Ala His Ala Ala Gly lie Ala Gly Ser Leu Thr Asn Lys Lys Val Asn 135 140 145 150

AGA ATT ACT GGC CTC GAT CCA GCT GGA CCT AAC TTT GAG TAT GCA GAA 753 Arg lie Thr Gly Leu Asp Pro Ala Gly Pro Asn Phe Glu Tyr Ala Glu 155 160 165

GCC CCG AGT CGT CTT TCT CCT GAT GAT GCA GAT TTT GTA GAC GTC TTA 801 Ala Pro Ser Arg Leu Ser Pro Asp Asp Ala Asp Phe Val Asp Val Leu 170 175 180 CAC ACA TTC ACC AGA GGG TCC CCT GGT CGA AGC ATT GGA ATC CAG AAA 849 His Thr Phe Thr Arg Gly Ser Pro Gly Arg Ser lie Gly lie Gin Lys 185 190 195

CCA GTT GGG CAT GTT GAC ATT TAC CCG AAT GGA GGT ACT TTT CAG CCA 897 Pro Val Gly His Val Asp lie Tyr Pro Asn Gly Gly Thr Phe Gin Pro 200 205 210

GGA TGT AAC ATT GGA GAA GCT ATC CGC GTG ATT GCA GAG AGA GGA CTT 945 Gly Cys Asn lie Gly Glu Ala lie Arg Val lie Ala Glu Arg Gly Leu 215 220 225 230

GGA GAT GTG GAC CAG CTA GTG AAG TGC TCC CAC GAG CGC TCC ATT CAT 993

Gly Asp Val Asp Gin Leu Val Lys Cys Ser His Glu Arg Ser lie His

235 240 245

CTC TTC ATC GAC TCT CTG TTG AAT GAA GAA AAT CCA AGT AAG GCC TAC 1041

Leu Phe lie Asp Ser Leu Leu Asn Glu Glu Asn Pro Ser Lys Ala Tyr

250 255 260 AGG TGC AGT TCC AAG GAA GCC TTT GAG AAA GGG CTC TGC TTG AGT TGT 1089 Arg Cys Ser Ser Lys Glu Ala Phe Glu Lys Gly Leu Cys Leu Ser Cys 265 270 275 AGA AAG AAC CGC TGC AAC AAT CTG GGC TAT GAG ATC AAT AAA GTC AGA 1137 Arg Lys Asn Arg Cys Asn Asn Leu Gly Tyr Glu lie Asn Lys Val Arg 280 285 290

GCC AAA AGA AGC AGC AAA ATG TAC CTG AAG ACT CGT TCT CAG ATG CCC 1185 Ala Lys Arg Ser Ser Lys Met Tyr Leu Lys Thr Arg Ser Gin Met Pro 295 300 305 310 TAC AAA GTC TTC CAT TAC CAA GTA AAG ATT CAT TTT TCT GGG ACT GAG 1233 Tyr Lys Val Phe His Tyr Gin Val Lys lie His Phe Ser Gly Thr Glu 315 320 325

AGT GAA ACC CAT ACC AAT CAG GCC TTT GAG ATT TCT CTG TAT GGC ACC 1281 Ser Glu Thr His Thr Asn Gin Ala Phe Glu lie Ser Leu Tyr Gly Thr

330 335 340

GTG GCC GAG AGT GAG AAC ATC CCA TTC ACT CTG CCT GAA GTT TCC ACA 1329 Val Ala Glu Ser Glu Asn lie Pro Phe Thr Leu Pro Glu Val Ser Thr 345 350 355

AAT AAG ACC TAC TCC TTC CTA ATT TAC ACA GAG GTA GAT ATT GGA GAA 1377 Asn Lys Thr Tyr Ser Phe Leu lie Tyr Thr Glu Val Asp lie Gly Glu 360 365 370

CTA CTC ATG TTG AAG CTC AAA TGG AAG AGT GAT TCA TAC TTT AGC TGG 1425 Leu Leu Met Leu Lys Leu Lys Trp Lys Ser Asp Ser Tyr Phe Ser Trp 375 380 385 390 TCA GAC TGG TGG AGC AGT CCC GGC TTC GCC ATT CAG AAG ATC AGA GTA 1473 Ser Asp Trp Trp Ser Ser Pro Gly Phe Ala lie Gin Lys lie Arg Val 395 400 405

AAA GCA GGA GAG ACT CAG AAA AAG GTG ATC TTC TGT TCT AGG GAG AAA 1521 Lys Ala Gly Glu Thr Gin Lys Lys Val lie Phe Cys Ser Arg Glu Lys

410 415 420

GTG TCT CAT TTG CAG AAA GGA AAG GCA CCT GCG GTA TTT GTG AAA TGC 1569 Val Ser His Leu Gin Lys Gly Lys Ala Pro Ala Val Phe Val Lys Cys 425 430 435

CAT GAC AAG TCT CTG AAT AAG AAG TCA GGC TGA AACTGGGCGA ATCTACAGAA 1622 His Asp Lys Ser Leu Asn Lys Lys Ser Gly * 440 445

CAAAGAACGG CATGTGAATT CTGTGAAGAA TGAAGTGGAG GAAGTAACTT TTACAAAACA 1682

TACCCAGTGT TTGGGGTGTT TCAAAAGTGG ATTTTCCTGA ATATTAATCC CAGCCCTACC 1742 CTTGTTAGTT ATTTTAGGAG ACAGTCTCAA GCACTAAAAA GTGGCTAATT CAATTTATGG 1802

GGTATAGTGG CCAAATAGCA CATCCTCCAA CGTTAAAAGA CAGTGGATCA TGAAAAGTGC 1862

TGTTTTGTCC TTTGAGAAAG AAATAATTGT TTGAGCGCAG AGTAAAATAA GGCTCCTTCA 1922 TGTGGCGTAT TGGGCCATAG CCTATAATTG GTTAGAACCT CCTATTTTAA TTGGAATTCT 1982

GGATCTTTCG GACTGAGGCC TTCTCAAACT TTACTCTAAG TCTCCAAGAA TACAGAAAAT 2042

GCTTTTCCGC GGCACGAATC AGACTCATCT ACACAGCAGT ATGAATGATG TTTTAGAATG 2102

ATTCCCTCTT GCTATTGGAA TGTGGTCCAG ACGTCAACCA GGAACATGTA ACTTGGAGAG 2162

GGACGAAGAA AGGGTCTGAT AAACACAGAG GTTTTAAACA GTCCCTACCA TTGGCCTGCA 2222

TCATGACAAA GTTACAAATT CAAGGAGATA TAAAATCTAG ATCAATTAAT TCTTAATAGG 2282

CTTTATCGTT TATTGCTTAA TCCCTCTCTC CCCCTTCTTT TTTGTCTCAA GATTATATTA 2342 TAATAATGTT CTCTGGGTAG GTGTTGAAAA TGAGCCTGTA ATCCTCAGCT GACACATAAT 2402

TTGAATGGTG CAGAAAAAAA AAAGATACCG TAATTTTATT ATTAGATTCT CCAAATGATT 2462

TTCATCAATT TAAAATCATT CAATATCTGA CAGTTACTCT TCAGTTTTAG GCTTACCTTG 2522

GTCCATGCTC AGTTGTACTT CCAGTGCGTC TCTTTTGTTC CTGGCTTTGA CATGAAAAGA 2582

TAGGTTTGAG TTCAAATTTT GCATTGTGTG AGCTTCTACA GATTTTAGAC AAGGACCGTT 2642 TTTACTAAGT AAAAGGGTGG AGAGGTTCCT GGGGTGGATT CCTAAGCAGT GCTTGTAAAC 2702

CATCGCGTGC AATGAGCCAG ATGGAGTACC ATGAGGGTTG TTATTTGTTG TTTTTAACAA 2762

CTAATCAAGA GTGAGTGAAC AACTATTTAT AAACTAGATC TCCTATTTTT CAGAATGCTC 2822

TTCTACGTAT AAATATGAAA TGATAAAGAT GTCAAATATC TCAGAGGCTA TAGCTGGGAA 2882

CCCGACTGTG AAAGTATGTG ATATCTGAAC ACATACTAGA AAGCTCTGCA TGTGTGTTGT 2942 CCTTCAGCAT AATTCGGAAG GGAAAACAGT CGATCAAGGG ATGTATTGGA ACATGTCGGA 3002

GTAGAAATTG TTCCTGATGT GCCAGAACTT CGACCCTTTC TCTGAGAGAG ATGATCGTGC 3062

CTATAAATAG TAGGACCAAT GTTGTGATTA ACATCATCAG GCTTGGAATG AATTCTCTCT 3122

AAAAATAAAA TGATGTATGA TTTGTTGTTG GCATCCCCTT TATTAATTCA TTAAATTTCT 3182

GGATTTGGGT TGTGACCCAG GGTGCATTAA CTTAAAAGAT TCACTAAAGC AGCACATAGC 3242 ACTGGGAACT CTGGCTCCGA AAAACTTTGT TATATATATC AAGGATGTTC TGGCTTTACA 3302

TTTTATTTAT TAGCTGTAAA TACATGTGTG GATGTGTAAA TGGAGCTTGT ACATATTGGA 3362

AAGGTCATTG TGGCTATCTG CATTTATAAA TGTGTGGTGC TAACTGTATG TGTCTTTATC 3422

AGTGATGGTC TCACAGAGCC AACTCACTCT TATGAAATGG GCTTTAACAA AACAAGAAAG 3482

AAACGTACTT AACTGTGTGA AGAAATGGAA TCAGCTTTTA ATAAAATTGA CAACATTTTA 3542 TTACCAC 3549 (2) INFORMATION FOR SEQ ID NO: 6:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 476 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:

Met Glu Ser Lys Ala Leu Leu Val Leu Thr Leu Ala Val Trp Leu Gin -27 -25 -20 -15

Ser Leu Thr Ala Ser Arg Gly Gly Val Ala Ala Ala Asp Gin Arg Arg -10 -5 1 5

Asp Phe lie Asp lie Glu Ser Lys Phe Ala Leu Arg Thr Pro Glu Asp 10 15 20

Thr Ala Glu Asp Thr Cys His Leu lie Pro Gly Val Ala Glu Ser Val

25 30 35 Ala Thr Cys His Phe Asn His Ser Ser Lys Thr Phe Met Val lie His

40 45 50

Gly Trp Thr Val Thr Gly Met Tyr Glu Ser Trp Val Pro Lys Leu Val 55 60 65

Ala Ala Leu Tyr Lys Arg Glu Pro Asp Ser Asn Val lie Val Val Asp 70 75 80 85

Trp Leu Ser Arg Ala Gin Glu His Tyr Pro Val Ser Ala Gly Tyr Thr 90 95 100

Lys Leu Val Gly Gin Asp Val Ala Arg Phe lie Asn Trp Met Glu Glu 105 110 115 Glu Phe Asn Tyr Pro Leu Asp Asn Val His Leu Leu Gly Tyr Ser Leu 120 125 130

Gly Ala His Ala Ala Gly lie Ala Gly Ser Leu Thr Asn Lys Lys Val 135 140 145

Asn Arg lie Thr Gly Leu Asp Pro Ala Gly Pro Asn Phe Glu Tyr Ala 150 155 160 165

Glu Ala Pro Ser Arg Leu Ser Pro Asp Asp Ala Asp Phe Val Asp Val 170 175 180

Leu His Thr Phe Thr Arg Gly Ser Pro Gly Arg Ser lie Gly lie Gin

185 190 195 Lys Pro Val Gly His Val Asp lie Tyr Pro Asn Gly Gly Thr Phe Gin 200 205 210

Pro Gly Cys Asn lie Gly Glu Ala lie Arg Val lie Ala Glu Arg Gly 215 220 225

Leu Gly Asp Val Asp Gin Leu Val Lys Cys Ser His Glu Arg Ser lie 230 235 240 245

His Leu Phe lie Asp Ser Leu Leu Asn Glu Glu Asn Pro Ser Lys Ala 250 255 260

Tyr Arg Cys Ser Ser Lys Glu Ala Phe Glu Lys Gly Leu Cys Leu Ser 265 270 275

Cys Arg Lys Asn Arg Cys Asn Asn Leu Gly Tyr Glu lie Asn Lys Val

280 285 290 Arg Ala Lys Arg Ser Ser Lys Met Tyr Leu Lys Thr Arg Ser Gin Met

295 300 305

Pro Tyr Lys Val Phe His Tyr Gin Val Lys lie His Phe Ser Gly Thr

310 315 320 325

Glu Ser Glu Thr His Thr Asn Gin Ala Phe Glu lie Ser Leu Tyr Gly

330 335 340

Thr Val Ala Glu Ser Glu Asn lie Pro Phe Thr Leu Pro Glu Val Ser 345 350 355

Thr Asn Lys Thr Tyr Ser Phe Leu lie Tyr Thr Glu Val Asp lie Gly 360 365 370 Glu Leu Leu Met Leu Lys Leu Lys Trp Lys Ser Asp Ser Tyr Phe Ser 375 380 385

Trp Ser Asp Trp Trp Ser Ser Pro Gly Phe Ala lie Gin Lys lie Arg 390 395 400 405

Val Lys Ala Gly Glu Thr Gin Lys Lys Val lie Phe Cys Ser Arg Glu 410 415 420

Lys Val Ser His Leu Gin Lys Gly Lys Ala Pro Ala Val Phe Val Lys 425 430 435

Cys His Asp Lys Ser Leu Asn Lys Lys Ser Gly * 440 445 (2) INFORMATION FOR SEQ ID NO : 7 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 4020 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:

(A) ORGANISM: Homo sapiens

(ix) FEATURE:

(A) NAME/KEY: sig_peptide (B) LOCATION: 1..109

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 23..3943

(ix) FEATURE:

(A) NAME/KEY: mat_peptide

(B) LOCATION: 110..3940

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 7 :

GCCGAGCACC GCGCACCGCG TC ATG GGG GCC GCC TCG GGC CGC CGG GGG CCG 52

Met Gly Ala Ala Ser Gly Arg Arg Gly Pro -29 -25 -20

GGG CTG CTG CTG CCG CTG CCG CTG CTG TTG CTG CTG CCG CCG CAG CCC 100 Gly Leu Leu Leu Pro Leu Pro Leu Leu Leu Leu Leu Pro Pro Gin Pro -15 -10 -5

GCC CTG GCG TTG GAC CCC GGG CTG CAG CCC GGC AAC TTT TCT GCT GAC 148 Ala Leu Ala Leu Asp Pro Gly Leu Gin Pro Gly Asn Phe Ser Ala Asp 1 5 10 GAG GCC GGG GCG CAG CTC TTC GCG CAG AGC TAC AAC TCC AGC GCC GAA 196 Glu Ala Gly Ala Gin Leu Phe Ala Gin Ser Tyr Asn Ser Ser Ala Glu 15 20 25

CAG GTG CTG TTC CAG AGC GTG GCC GCC AGC TGG GCG CAC GAC ACC AAC 244 Gin Val Leu Phe Gin Ser Val Ala Ala Ser Trp Ala His Asp Thr Asn 30 35 40 45

ATC ACC GCG GAG AAT GCA AGG CGC CAG GAG GAA GCA GCC CTG CTC AGC 292 lie Thr Ala Glu Asn Ala Arg Arg Gin Glu Glu Ala Ala Leu Leu Ser 50 55 60

CAG GAG TTT GCG GAG GCC TGG GGC CAG AAG GCC AAG GAG CTG TAT GAA 340 Gin Glu Phe Ala Glu Ala Trp Gly Gin Lys Ala Lys Glu Leu Tyr Glu 65 70 75

CCG ATC TGG CAG AAC TTC ACG GAC CCG CAG CTG CGC AGG ATC ATC GGA 388 Pro lie Trp Gin Asn Phe Thr Asp Pro Gin Leu Arg Arg lie lie Gly 80 85 90 GCT GTG CGA ACC CTG GGC TCT GCC AAC CTG CCC CTG GCT AAG CGG CAG 436 Ala Val Arg Thr Leu Gly Ser Ala Asn Leu Pro Leu Ala Lys Arg Gin 95 100 105

CAG TAC AAC GCC CTG CTA AGC AAC ATG AGC AGG ATC TAC TCC ACC GCC 484 Gin Tyr Asn Ala Leu Leu Ser Asn Met Ser Arg He Tyr Ser Thr Ala 110 115 120 125 AAG GTC TGC CTC CCC AAC AAG ACT GCC ACC TGC TGG TCC CTG GAC CCA 532 Lys Val Cys Leu Pro Asn Lys Thr Ala Thr Cys Trp Ser Leu Asp Pro 130 135 140

GAT CTC ACC AAC ATC CTG GCT TCC TCG CGA AGC TAC GCC ATG CTC CTG 580 Asp Leu Thr Asn He Leu Ala Ser Ser Arg Ser Tyr Ala Met Leu Leu

145 150 155

TTT GCC TGG GAG GGC TGG CAC AAC GCT GCG GGC ATC CCG CTG AAA CCG 628 Phe Ala Trp Glu Gly Trp His Asn Ala Ala Gly He Pro Leu Lys Pro 160 165 170

CTG TAC GAG GAT TTC ACT GCC CTC AGC AAT GAA GCC TAC AAG CAG GAC 676

Leu Tyr Glu Asp Phe Thr Ala Leu Ser Asn Glu Ala Tyr Lys Gin Asp

175 180 185

GGC TTC ACA GAC ACG GGG GCC TAC TGG CGC TCC TGG TAC AAC TCC CCC 724

Gly Phe Thr Asp Thr Gly Ala Tyr Trp Arg Ser Trp Tyr Asn Ser Pro 190 195 200 205 ACC TTC GAG GAC GAT CTG GAA CAC CTC TAC CAA CAG CTA GAG CCC CTC 772 Thr Phe Glu Asp Asp Leu Glu His Leu Tyr Gin Gin Leu Glu Pro Leu 210 215 220

TAC CTG AAC CTC CAT GCC TTC GTC CGC CGC GCA CTG CAT CGC CGA TAC 820 Tyr Leu Asn Leu His Ala Phe Val Arg Arg Ala Leu His Arg Arg Tyr

225 230 235

GGA GAC AGA TAC ATC AAC CTC AGG GGA CCC ATC CCT GCT CAT CTG CTG 868 Gly Asp Arg Tyr He Asn Leu Arg Gly Pro He Pro Ala His Leu Leu 240 245 250

GGA GAC ATG TGG GCC CAG AGC TGG GAA AAC ATC TAC GAC ATG GTG GTG 916 Gly Asp Met Trp Ala Gin Ser Trp Glu Asn He Tyr Asp Met Val Val 255 260 265

CCT TTC CCA GAC AAG CCC AAC CTC GAT GTC ACC AGT ACT ATG CTG CAG 964 Pro Phe Pro Asp Lys Pro Asn Leu Asp Val Thr Ser Thr Met Leu Gin 270 275 280 285 CAG GGC TGG AAC GCC ACG CAC ATG TTC CGG GTG GCA GAG GAG TTC TTC 1012 Gin Gly Trp Asn Ala Thr His Met Phe Arg Val Ala Glu Glu Phe Phe 290 295 300 ACC TCC CTG GAG CTC TCC CCC ATG CCT CCC GAG TTC TGG GAA GGG TCG 1060 Thr Ser Leu Glu Leu Ser Pro Met Pro Pro Glu Phe Trp Glu Gly Ser 305 310 315

ATG CTG GAG AAG CCG GCC GAC GGG CGG GAA GTG GTG TGC CAC GCC TCG 1108 Met Leu Glu Lys Pro Ala Asp Gly Arg Glu Val Val Cys His Ala Ser 320 325 330 GCT TGG GAC TTC TAC AAC AGG AAA GAC TTC AGG ATC AAG CAG TGC ACA 1156 Ala Trp Asp Phe Tyr Asn Arg Lys Asp Phe Arg He Lys Gin Cys Thr 335 340 345

CGG GTC ACG ATG GAC CAG CTC TCC ACA GTG CAC CAT GAG ATG GGC CAT 1204 Arg Val Thr Met Asp Gin Leu Ser Thr Val His His Glu Met Gly His 350 355 360 365

ATA CAG TAC TAC CTG CAG TAC AAG GAT CTG CCC GTC TCC CTG CGT CGG 1252 He Gin Tyr Tyr Leu Gin Tyr Lys Asp Leu Pro Val Ser Leu Arg Arg 370 375 380

GGG GCC AAC CCC GGC TTC CAT GAG GCC ATT GGG GAC GTG CTG GCG CTC 1300

Gly Ala Asn Pro Gly Phe His Glu Ala He Gly Asp Val Leu Ala Leu 385 390 395

TCG GTC TCC ACT CCT GAA CAT CTG CAC AAA ATC GGC CTG CTG GAC CGT 1348

Ser Val Ser Thr Pro Glu His Leu His Lys He Gly Leu Leu Asp Arg 400 405 410 GTC ACC AAT GAC ACG GAA AGT GAC ATC AAT TAC TTG CTA AAA ATG GCA 1396 Val Thr Asn Asp Thr Glu Ser Asp He Asn Tyr Leu Leu Lys Met Ala 415 420 425

CTG GAA AAA ATT GCC TTC CTG CCC TTT GGC TAC TTG GTG GAC CAG TGG 1444 Leu Glu Lys He Ala Phe Leu Pro Phe Gly Tyr Leu Val Asp Gin Trp 430 435 440 445

CGC TGG GGG GTC TTT AGT GGG CGT ACC CCC CCT TCC CGC TAC AAC TTC 1492 Arg Trp Gly Val Phe Ser Gly Arg Thr Pro Pro Ser Arg Tyr Asn Phe 450 455 460

GAC TGG TGG TAT CTT CGA ACC AAG TAT CAG GGG ATC TGT CCT CCT GTT 1540 Asp Trp Trp Tyr Leu Arg Thr Lys Tyr Gin Gly He Cys Pro Pro Val 465 470 475

ACC CGA AAC GAA ACC CAC TTT GAT GCT GGA GCT AAG TTT CAT GTT CCA 1588 Thr Arg Asn Glu Thr His Phe Asp Ala Gly Ala Lys Phe His Val Pro 480 485 490 AAT GTG ACA CCA TAC ATC AGG TAC TTT GTG AGT TTT GTC CTG CAG TTC 1636 Asn Val Thr Pro Tyr He Arg Tyr Phe Val Ser Phe Val Leu Gin Phe 495 500 505 CAG TTC CAT GAA GCC CTG TGC AAG GAG GCA GGC TAT GAG GGC CCA CTG 1684 Gin Phe His Glu Ala Leu Cys Lys Glu Ala Gly Tyr Glu Gly Pro Leu 510 515 520 525

CAC CAG TGT GAC ATC TAC CGG TCC ACC AAG GCA GGG GCC AAG CTC CGG 1732 His Gin Cys Asp He Tyr Arg Ser Thr Lys Ala Gly Ala Lys Leu Arg 530 535 540 AAG GTG CTG CAG GCT GGC TCC TCC AGG CCC TGG CAG GAG GTG CTG AAG 1780 Lys Val Leu Gin Ala Gly Ser Ser Arg Pro Trp Gin Glu Val Leu Lys 545 550 555

GAC ATG GTC GGC TTA GAT GCC CTG GAT GCC CAG CCG CTG CTC AAG TAC 1828 Asp Met Val Gly Leu Asp Ala Leu Asp Ala Gin Pro Leu Leu Lys Tyr 560 565 570

TTC CAG CCA GTC ACC CAG TGG CTG CAG GAG CAG AAC CAG CAG AAC GGC 1876 Phe Gin Pro Val Thr Gin Trp Leu Gin Glu Gin Asn Gin Gin Asn Gly 575 580 585

GAG GTC CTG GGC TGG CCC GAG TAC CAG TGG CAC CCG CCG TTG CCT GAC 1924 Glu Val Leu Gly Trp Pro Glu Tyr Gin Trp His Pro Pro Leu Pro Asp 590 595 600 605

AAC TAC CCG GAG GGC ATA GAC CTG GTG ACT GAT GAG GCT GAG GCC AGC 1972 Asn Tyr Pro Glu Gly He Asp Leu Val Thr Asp Glu Ala Glu Ala Ser 610 615 620 AAG TTT GTG GAG GAA TAT GAC CGG ACA TCC CAG GTG GTG TGG AAC GAG 2020 Lys Phe Val Glu Glu Tyr Asp Arg Thr Ser Gin Val Val Trp Asn Glu 625 630 635

TAT GCC GAG GCC AAC TGG AAC TAC AAC ACC AAC ATC ACC ACA GAG ACC 2068 Tyr Ala Glu Ala Asn Trp Asn Tyr Asn Thr Asn He Thr Thr Glu Thr 640 645 650

AGC AAG ATT CTG CTG CAG AAG AAC ATG CAA ATA GCC AAC CAC ACC CTG 2116 Ser Lys He Leu Leu Gin Lys Asn Met Gin He Ala Asn His Thr Leu 655 660 665

AAG TAC GGC ACC CAG GCC AGG AAG TTT GAT GTG AAC CAG TTG CAG AAC 2164 Lys Tyr Gly Thr Gin Ala Arg Lys Phe Asp Val Asn Gin Leu Gin Asn 670 675 680 685

ACC ACT ATC AAG CGG ATC ATA AAG AAG GTT CAG GAC CTA GAA CGG GCA 2212 Thr Thr He Lys Arg He He Lys Lys Val Gin Asp Leu Glu Arg Ala 690 695 700 GCG CTG CCT GCC CAG GAG CTG GAG GAG TAC AAC AAG ATC CTG TTG GAT 2260 Ala Leu Pro Ala Gin Glu Leu Glu Glu Tyr Asn Lys He Leu Leu Asp 705 710 715 ATG GAA ACC ACC TAC AGC GTG GCC ACT GTG TGC CAC CCG AAT GGC AGC 2308

Met Glu Thr Thr Tyr Ser Val Ala Thr Val Cys His Pro Asn Gly Ser 720 725 730

TGC CTG CAG CTC GAG CCA GAT CTG ACG AAT GTG ATG GCC ACA TCC CGG 2356

Cys Leu Gin Leu Glu Pro Asp Leu Thr Asn Val Met Ala Thr Ser Arg 735 740 745 AAA TAT GAA GAC CTG TTA TGG GCA TGG GAG GGC TGG CGA GAC AAG GCG 2404 Lys Tyr Glu Asp Leu Leu Trp Ala Trp Glu Gly Trp Arg Asp Lys Ala 750 755 760 765

GGG AGA GCC ATC CTC CAG TTT TAC CCG AAA TAC GTG GAA CTC ATC AAC 2452 Gly Arg Ala He Leu Gin Phe Tyr Pro Lys Tyr Val Glu Leu He Asn

770 775 780

CAG GCT GCC CGG CTC AAT GGC TAT GTA GAT GCA GGG GAC TCG TGG AGG 2500 Gin Ala Ala Arg Leu Asn Gly Tyr Val Asp Ala Gly Asp Ser Trp Arg 785 790 795

TCT ATG TAC GAG ACA CCA TCC CTG GAG CAA GAC CTG GAG CGG CTC TTC 2548

Ser Met Tyr Glu Thr Pro Ser Leu Glu Gin Asp Leu Glu Arg Leu Phe 800 805 810

CAG GAG CTG CAG CCA CTC TAC CTC AAC CTG CAT GCC TAC GTG CGC CGG 2596

Gin Glu Leu Gin Pro Leu Tyr Leu Asn Leu His Ala Tyr Val Arg Arg

815 820 825 GCC CTG CAC CGT CAC TAC GGG GCC CAG CAC ATC AAC CTG GAG GGG CCC 2644 Ala Leu His Arg His Tyr Gly Ala Gin His He Asn Leu Glu Gly Pro 830 835 840 845

ATT CCT GCT CAC CTG CTG GGG AAC ATG TGG GCG CAG ACC TGG TCC AAC 2692 He Pro Ala His Leu Leu Gly Asn Met Trp Ala Gin Thr Trp Ser Asn

850 855 860

ATC TAT GAC TTG GTG GTG CCC TTC CCT TCA GCC CCC TCG ATG GAC ACC 2740 He Tyr Asp Leu Val Val Pro Phe Pro Ser Ala Pro Ser Met Asp Thr 865 870 875

ACA GAG GCT ATG CTA AAG CAG GGC TGG ACG CCC AGG AGG ATG TTT AAG 2788 Thr Glu Ala Met Leu Lys Gin Gly Trp Thr Pro Arg Arg Met Phe Lys 880 885 890

GAG GCT GAT GAT TTC TTC ACC TCC CTG GGG CTG CTG CCC GTG CCT CCT 2836 Glu Ala Asp Asp Phe Phe Thr Ser Leu Gly Leu Leu Pro Val Pro Pro 895 900 905 GAG TTC TGG AAC AAG TCG ATG CTG GAG AAG CCA ACC GAC GGG CGG GAG 2884 Glu Phe Trp Asn Lys Ser Met Leu Glu Lys Pro Thr Asp Gly Arg Glu 910 915 920 925 GTG GTC TGC CAC GCC TCG GCC TGG GAC TTC TAC AAC GGC AAG GAC TTC 2932

Val Val Cys His Ala Ser Ala Trp Asp Phe Tyr Asn Gly Lys Asp Phe

930 935 940

CGG ATC AAG CAG TGC ACC ACC GTG AAC TTG GAG GAC CTG GTG GTG GCC 2980

Arg He Lys Gin Cys Thr Thr Val Asn Leu Glu Asp Leu Val Val Ala

945 950 955 CAC CAC GAA ATG GGC CAC ATC CAG TAT TTC ATG CAG TAC AAA GAC TTA 3028 His His Glu Met Gly His He Gin Tyr Phe Met Gin Tyr Lys Asp Leu 960 965 970

CCT GTG GCC TTG AGG GAG GGT GCC AAC CCC GGC TTC CAT GAG GCC ATT 3076 Pro Val Ala Leu Arg Glu Gly Ala Asn Pro Gly Phe His Glu Ala He 975 980 985

GGG GAC GTG CTA GCC CTC TCA GTG TCT ACG CCC AAG CAC CTG CAC AGT 3124 Gly Asp Val Leu Ala Leu Ser Val Ser Thr Pro Lys His Leu His Ser 990 995 1000 1005

CTC AAC CTG CTG AGC AGT GAG GGT GGC AGC GAC GAG CAT GAC ATC AAC 3172 Leu Asn Leu Leu Ser Ser Glu Gly Gly Ser Asp Glu His Asp He Asn 1010 1015 1020

TTT CTG ATG AAG ATG GCC CTT GAC AAG ATC GCC TTT ATC CCC TTC AGC 3220 Phe Leu Met Lys Met Ala Leu Asp Lys He Ala Phe He Pro Phe Ser 1025 1030 1035 TAC CTC GTC GAT CAG TGG CGC TGG AGG GTA TTT GAT GGA AGC ATC ACC 3268 Tyr Leu Val Asp Gin Trp Arg Trp Arg Val Phe Asp Gly Ser He Thr 1040 1045 1050

AAG GAG AAC TAT AAC CAG GAG TGG TGG AGC CTC AGG CTG AAG TAC CAG 3316 Lys Glu Asn Tyr Asn Gin Glu Trp Trp Ser Leu Arg Leu Lys Tyr Gin 1055 1060 1065

GGC CTC TGC CCC CCA GTG CCC AGG ACT CAA GGT GAC TTT GAC CCA GGG 3364 Gly Leu Cys Pro Pro Val Pro Arg Thr Gin Gly Asp Phe Asp Pro Gly 1070 1075 1080 1085

GCC AAG TTC CAC ATT CCT TCT AGC GTG CCT TAC ATC AGG TAC TTT GTC 3412

Ala Lys Phe His He Pro Ser Ser Val Pro Tyr He Arg Tyr Phe Val 1090 1095 1100

AGC TTC ATC ATC CAG TTC CAG TTC CAC GAG GCA CTG TGC CAG GCA GCT 3460

Ser Phe He He Gin Phe Gin Phe His Glu Ala Leu Cys Gin Ala Ala 1105 1110 1115 GGC CAC ACG GGC CCC CTG CAC AAG TGT GAC ATC TAC CAG TCC AAG GAG 3508 Gly His Thr Gly Pro Leu His Lys Cys Asp He Tyr Gin Ser Lys Glu 1120 1125 1130 GCC GGG CAG CGC CTG GCG ACC GCC ATG AAG CTG GGC TTC AGT AGG CCG 3556 Ala Gly Gin Arg Leu Ala Thr Ala Met Lys Leu Gly Phe Ser Arg Pro 1135 1140 1145

TGG CCG GAA GCC ATG CAG CTG ATC ACG GGC CAG CCC AAC ATG AGC GCC 3604 Trp Pro Glu Ala Met Gin Leu He Thr Gly Gin Pro Asn Met Ser Ala 1150 1155 1160 1165 TCG GCC ATG TTG AGC TAC TTC AAG CCG CTG CTG GAC TGG CTC CGC ACG 3652 Ser Ala Met Leu Ser Tyr Phe Lys Pro Leu Leu Asp Trp Leu Arg Thr 1170 1175 1180

GAG AAC GAG CTG CAT GGG GAG AAG CTG GGC TGG CCG CAG TAC AAC TGG 3700 Glu Asn Glu Leu His Gly Glu Lys Leu Gly Trp Pro Gin Tyr Asn Trp

1185 1190 1195

ACG CCG AAC TCC GCT CGC TCA GAA GGG CCC CTC CCA GAC AGC GGC CGC 3748 Thr Pro Asn Ser Ala Arg Ser Glu Gly Pro Leu Pro Asp Ser Gly Arg 1200 1205 1210

GTC AGC TTC CTG GGC CTG GAC CTG GAT GCG CAG CAG GCC CGC GTG GGC 3796 Val Ser Phe Leu Gly Leu Asp Leu Asp Ala Gin Gin Ala Arg Val Gly 1215 1220 1225

CAG TGG CTG CTG CTC TTC CTG GGC ATC GCC CTG CTG GTA GCC ACC CTG 3844 Gin Trp Leu Leu Leu Phe Leu Gly He Ala Leu Leu Val Ala Thr Leu 1230 1235 1240 1245 GGC CTC AGC CAG CGG CTC TTC AGC ATC CGC CAC CGC AGC CTC CAC CGG 3892 Gly Leu Ser Gin Arg Leu Phe Ser He Arg His Arg Ser Leu His Arg 1250 1255 1260

CAC TCC CAC GGG CCC CAG TTC GGC TCC GAG GTG GAG CTG AGA CAC TCC 3940 His Ser His Gly Pro Gin Phe Gly Ser Glu Val Glu Leu Arg His Ser

1265 1270 1275

TGA GGTGACCCGG CTGGGTCGGC CCTGCCCAAG GGCCTCCCAC CAGAGACTGG 3993

GATGGGAACA CTGGTGGGCA GCTGAGG 4020

(2) INFORMATION FOR SEQ ID NO: 8:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1307 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 8 : Met Gly Ala Ala Ser Gly Arg Arg Gly Pro Gly Leu Leu Leu Pro Leu

-29 -25 -20 -15

Pro Leu Leu Leu Leu Leu Pro Pro Gin Pro Ala Leu Ala Leu Asp Pro -10 -5 1

Gly Leu Gin Pro Gly Asn Phe Ser Ala Asp Glu Ala Gly Ala Gin Leu

5 10 15 Phe Ala Gin Ser Tyr Asn Ser Ser Ala Glu Gin Val Leu Phe Gin Ser

20 25 30 35

Val Ala Ala Ser Trp Ala His Asp Thr Asn He Thr Ala Glu Asn Ala 40 45 50

Arg Arg Gin Glu Glu Ala Ala Leu Leu Ser Gin Glu Phe Ala Glu Ala 55 60 65

Trp Gly Gin Lys Ala Lys Glu Leu Tyr Glu Pro He Trp Gin Asn Phe 70 75 80

Thr Asp Pro Gin Leu Arg Arg He He Gly Ala Val Arg Thr Leu Gly

85 90 95 Ser Ala Asn Leu Pro Leu Ala Lys Arg Gin Gin Tyr Asn Ala Leu Leu

100 105 110 115

Ser Asn Met Ser Arg He Tyr Ser Thr Ala Lys Val Cys Leu Pro Asn 120 125 130

Lys Thr Ala Thr Cys Trp Ser Leu Asp Pro Asp Leu Thr Asn He Leu 135 140 145

Ala Ser Ser Arg Ser Tyr Ala Met Leu Leu Phe Ala Trp Glu Gly Trp 150 155 160

His Asn Ala Ala Gly He Pro Leu Lys Pro Leu Tyr Glu Asp Phe Thr 165 170 175 Ala Leu Ser Asn Glu Ala Tyr Lys Gin Asp Gly Phe Thr Asp Thr Gly 180 185 190 195

Ala Tyr Trp Arg Ser Trp Tyr Asn Ser Pro Thr Phe Glu Asp Asp Leu 200 205 210

Glu His Leu Tyr Gin Gin Leu Glu Pro Leu Tyr Leu Asn Leu His Ala 215 220 225

Phe Val Arg Arg Ala Leu His Arg Arg Tyr Gly Asp Arg Tyr He Asn 230 235 240

Leu Arg Gly Pro He Pro Ala His Leu Leu Gly Asp Met Trp Ala Gin 245 250 255 Ser Trp Glu Asn He Tyr Asp Met Val Val Pro Phe Pro Asp Lys Pro 260 265 270 275

Asn Leu Asp Val Thr Ser Thr Met Leu Gin Gin Gly Trp Asn Ala Thr 280 285 290

His Met Phe Arg Val Ala Glu Glu Phe Phe Thr Ser Leu Glu Leu Ser 295 300 305

Pro Met Pro Pro Glu Phe Trp Glu Gly Ser Met Leu Glu Lys Pro Ala 310 315 320

Asp Gly Arg Glu Val Val Cys His Ala Ser Ala Trp Asp Phe Tyr Asn 325 330 335

Arg Lys Asp Phe Arg He Lys Gin Cys Thr Arg Val Thr Met Asp Gin 340 345 350 355 Leu Ser Thr Val His His Glu Met Gly His He Gin Tyr Tyr Leu Gin

360 365 370

Tyr Lys Asp Leu Pro Val Ser Leu Arg Arg Gly Ala Asn Pro Gly Phe 375 380 385

His Glu Ala He Gly Asp Val Leu Ala Leu Ser Val Ser Thr Pro Glu 390 395 400

His Leu His Lys He Gly Leu Leu Asp Arg Val Thr Asn Asp Thr Glu 405 410 415

Ser Asp He Asn Tyr Leu Leu Lys Met Ala Leu Glu Lys He Ala Phe 420 425 430 435 Leu Pro Phe Gly Tyr Leu Val Asp Gin Trp Arg Trp Gly Val Phe Ser

440 445 450

Gly Arg Thr Pro Pro Ser Arg Tyr Asn Phe Asp Trp Trp Tyr Leu Arg 455 460 465

Thr Lys Tyr Gin Gly He Cys Pro Pro Val Thr Arg Asn Glu Thr His 470 475 480

Phe Asp Ala Gly Ala Lys Phe His Val Pro Asn Val Thr Pro Tyr He 485 490 495

Arg Tyr Phe Val Ser Phe Val Leu Gin Phe Gin Phe His Glu Ala Leu 500 505 510 515 Cys Lys Glu Ala Gly Tyr Glu Gly Pro Leu His Gin Cys Asp He Tyr

520 525 530

Arg Ser Thr Lys Ala Gly Ala Lys Leu Arg Lys Val Leu Gin Ala Gly 535 540 545 Ser Ser Arg Pro Trp Gin Glu Val Leu Lys Asp Met Val Gly Leu Asp 550 555 560

Ala Leu Asp Ala Gin Pro Leu Leu Lys Tyr Phe Gin Pro Val Thr Gin 565 570 575

Trp Leu Gin Glu Gin Asn Gin Gin Asn Gly Glu Val Leu Gly Trp Pro 580 585 590 595 Glu Tyr Gin Trp His Pro Pro Leu Pro Asp Asn Tyr Pro Glu Gly He

600 605 610

Asp Leu Val Thr Asp Glu Ala Glu Ala Ser Lys Phe Val Glu Glu Tyr 615 620 625

Asp Arg Thr Ser Gin Val Val Trp Asn Glu Tyr Ala Glu Ala Asn Trp 630 635 640

Asn Tyr Asn Thr Asn He Thr Thr Glu Thr Ser Lys He Leu Leu Gin 645 650 655

Lys Asn Met Gin He Ala Asn His Thr Leu Lys Tyr Gly Thr Gin Ala 660 665 670 675 Arg Lys Phe Asp Val Asn Gin Leu Gin Asn Thr Thr He Lys Arg He

680 685 690

He Lys Lys Val Gin Asp Leu Glu Arg Ala Ala Leu Pro Ala Gin Glu 695 700 705

Leu Glu Glu Tyr Asn Lys He Leu Leu Asp Met Glu Thr Thr Tyr Ser 710 715 720

Val Ala Thr Val Cys His Pro Asn Gly Ser Cys Leu Gin Leu Glu Pro 725 730 735

Asp Leu Thr Asn Val Met Ala Thr Ser Arg Lys Tyr Glu Asp Leu Leu

740 745 750 755 Trp Ala Trp Glu Gly Trp Arg Asp Lys Ala Gly Arg Ala He Leu Gin

760 765 770

Phe Tyr Pro Lys Tyr Val Glu Leu He Asn Gin Ala Ala Arg Leu Asn 775 780 785

Gly Tyr Val Asp Ala Gly Asp Ser Trp Arg Ser Met Tyr Glu Thr Pro 790 795 800

Ser Leu Glu Gin Asp Leu Glu Arg Leu Phe Gin Glu Leu Gin Pro Leu 805 810 815

Tyr Leu Asn Leu His Ala Tyr Val Arg Arg Ala Leu His Arg His Tyr

820 825 830 835 Gly Ala Gin His He Asn Leu Glu Gly Pro He Pro Ala His Leu Leu 840 845 850

Gly Asn Met Trp Ala Gin Thr Trp Ser Asn He Tyr Asp Leu Val Val 855 860 865

Pro Phe Pro Ser Ala Pro Ser Met Asp Thr Thr Glu Ala Met Leu Lys 870 875 880

Gin Gly Trp Thr Pro Arg Arg Met Phe Lys Glu Ala Asp Asp Phe Phe 885 890 895

Thr Ser Leu Gly Leu Leu Pro Val Pro Pro Glu Phe Trp Asn Lys Ser 900 905 910 915

Met Leu Glu Lys Pro Thr Asp Gly Arg Glu Val Val Cys His Ala Ser 920 925 930 Ala Trp Asp Phe Tyr Asn Gly Lys Asp Phe Arg He Lys Gin Cys Thr

935 940 945

Thr Val Asn Leu Glu Asp Leu Val Val Ala His His Glu Met Gly His 950 955 960

He Gin Tyr Phe Met Gin Tyr Lys Asp Leu Pro Val Ala Leu Arg Glu 965 970 975

Gly Ala Asn Pro Gly Phe His Glu Ala He Gly Asp Val Leu Ala Leu 980 985 990 995

Ser Val Ser Thr Pro Lys His Leu His Ser Leu Asn Leu Leu Ser Ser 1000 1005 1010 Glu Gly Gly Ser Asp Glu His Asp He Asn Phe Leu Met Lys Met Ala

1015 1020 1025

Leu Asp Lys He Ala Phe He Pro Phe Ser Tyr Leu Val Asp Gin Trp 1030 1035 1040

Arg Trp Arg Val Phe Asp Gly Ser He Thr Lys Glu Asn Tyr Asn Gin 1045 1050 1055

Glu Trp Trp Ser Leu Arg Leu Lys Tyr Gin Gly Leu Cys Pro Pro Val 1060 1065 1070 1075

Pro Arg Thr Gin Gly Asp Phe Asp Pro Gly Ala Lys Phe His He Pro 1080 1085 1090 Ser Ser Val Pro Tyr He Arg Tyr Phe Val Ser Phe He He Gin Phe

1095 1100 1105

Gin Phe His Glu Ala Leu Cys Gin Ala Ala Gly His Thr Gly Pro Leu 1110 1115 1120 His Lys Cys Asp He Tyr Gin Ser Lys Glu Ala Gly Gin Arg Leu Ala 1125 1130 1135

Thr Ala Met Lys Leu Gly Phe Ser Arg Pro Trp Pro Glu Ala Met Gin 1140 1145 1150 1155

Leu He Thr Gly Gin Pro Asn Met Ser Ala Ser Ala Met Leu Ser Tyr 1160 1165 1170 Phe Lys Pro Leu Leu Asp Trp Leu Arg Thr Glu Asn Glu Leu His Gly

1175 1180 1185

Glu Lys Leu Gly Trp Pro Gin Tyr Asn Trp Thr Pro Asn Ser Ala Arg 1190 1195 1200

Ser Glu Gly Pro Leu Pro Asp Ser Gly Arg Val Ser Phe Leu Gly Leu 1205 1210 1215

Asp Leu Asp Ala Gin Gin Ala Arg Val Gly Gin Trp Leu Leu Leu Phe 1220 1225 1230 1235

Leu Gly He Ala Leu Leu Val Ala Thr Leu Gly Leu Ser Gin Arg Leu 1240 1245 1250 Phe Ser He Arg His Arg Ser Leu His Arg His Ser His Gly Pro Gin

1255 1260 1265

Phe Gly Ser Glu Val Glu Leu Arg His Ser * 1270 1275

(2) INFORMATION FOR SEQ ID NO : 9 :

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 25 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: YES (iv) ANTI-SENSE: YES

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 9 : CCGTTTGTGC AGGGCCTGGC TCTCT 25

(2) INFORMATION FOR SEQ ID NO: 10:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 25 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: YES (iv) ANTI-SENSE: YES

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10: CAGGGTGCTG TCCACACTGG ACCCC 25

(2) INFORMATION FOR SEQ ID NO: 11: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:

TTTAGGCCTG AAGTTTCCAC 20

(2) INFORMATION FOR SEQ ID NO : 12 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 19 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: CTCCCTAGAA GAGAAGATC 19 (2) INFORMATION FOR SEQ ID NO: 13:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 22 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: YES (iv) ANTI-SENSE: YES

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: TAGGAGGTTG AGGCACCTGT GC 22

(2) INFORMATION FOR SEQ ID NO : 14 :

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14: GTGGGTGAAT CACCTGAGGT C 21

(2) INFORMATION FOR SEQ ID NO: 15:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4604 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE:

(A) ORGANISM: Homo sapiens

(ix) FEATURE:

(A) NAME/KEY: mRNA (B) LOCATION: -1..4604

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 116..1399

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15:

GGAACAGCTT GTCCACCCGC CGGCCGGACC AGAAGCCTTT GGGTCTGAAG TGTCTGTGAG 60

ACCTCACAGA AGAGCACCCC TGGGCTCCAC TTACCTGCCC CCTGCTCCTT CAGGG ATG 118

Met

GAG GCA ATG GCG GCC AGC ACT TCC CTG CCT GAC CCT GGA GAC TTT GAC 166 Glu Ala Met Ala Ala Ser Thr Ser Leu Pro Asp Pro Gly Asp Phe Asp 1280 1285 1290 1295

CGG AAC GTG CCC CGG ATC TGT GGG GTG TGT GGA GAC CGA GCC ACT GGC 214 Arg Asn Val Pro Arg He Cys Gly Val Cys Gly Asp Arg Ala Thr Gly

1300 1305 1310

TTT CAC TTC AAT GCT ATG ACC TGT GAA GGC TGC AAA GGC TTC TTC AGG 262 Phe His Phe Asn Ala Met Thr Cys Glu Gly Cys Lys Gly Phe Phe Arg 1315 1320 1325

CGA AGC ATG AAG CGG AAG GCA CTA TTC ACC TGC CCC TTC AAC GGG GAC 310 Arg Ser Met Lys Arg Lys Ala Leu Phe Thr Cys Pro Phe Asn Gly Asp 1330 1335 1340

TGC CGC ATC ACC AAG GAC AAC CGA CGC CAC TGC CAG GCC TGC CGG CTC 358 Cys Arg He Thr Lys Asp Asn Arg Arg His Cys Gin Ala Cys Arg Leu 1345 1350 1355 AAA CGC TGT GTG GAC ATC GGC ATG ATG AAG GAG TTC ATT CTG ACA GAT 406 Lys Arg Cys Val Asp He Gly Met Met Lys Glu Phe He Leu Thr Asp 1360 1365 1370 1375

GAG GAA GTG CAG AGG AAG CGG GAG ATG ATC CTG AAG CGG AAG GAG GAG 454 Glu Glu Val Gin Arg Lys Arg Glu Met He Leu Lys Arg Lys Glu Glu

1380 1385 1390

GAG GCC TTG AAG GAC AGT CTG CGG CCC AAG CTG TCT GAG GAG CAG CAG 502 Glu Ala Leu Lys Asp Ser Leu Arg Pro Lys Leu Ser Glu Glu Gin Gin 1395 1400 1405 CGC ATC ATT GCC ATA CTG CTG GAC GCC CAC CAT AAG ACC TAC GAC CCC 550 Arg He He Ala He Leu Leu Asp Ala His His Lys Thr Tyr Asp Pro 1410 1415 1420 ACC TAC TCC GAC TTC TGC CAG TTC CGG CCT CCA GTT CGT GTG AAT GAT 598 Thr Tyr Ser Asp Phe Cys Gin Phe Arg Pro Pro Val Arg Val Asn Asp 1425 1430 1435

GGT GGA GGG AGC CAT CCT TCC AGG CCC AAC TCC AGA CAC ACT CCC AGC 646 Gly Gly Gly Ser His Pro Ser Arg Pro Asn Ser Arg His Thr Pro Ser 1440 1445 1450 1455

TTC TCT GGG GAC TCC TCC TCC TCC TGC TCA GAT CAC TGT ATC ACC TCT 694 Phe Ser Gly Asp Ser Ser Ser Ser Cys Ser Asp His Cys He Thr Ser 1460 1465 1470

TCA GAC ATG ATG GAC TCG TCC AGC TTC TCC AAT CTG GAT CTG AGT GAA 742 Ser Asp Met Met Asp Ser Ser Ser Phe Ser Asn Leu Asp Leu Ser Glu 1475 1480 1485

GAA GAT TCA GAT GAC CCT TCT GTG ACC CTA GAG CTG TCC CAG CTC TCC 790 Glu Asp Ser Asp Asp Pro Ser Val Thr Leu Glu Leu Ser Gin Leu Ser 1490 1495 1500 ATG CTG CCC CAC CTG GCT GAC CTG GTC AGT TAC AGC ATC CAA AAG GTC 838 Met Leu Pro His Leu Ala Asp Leu Val Ser Tyr Ser He Gin Lys Val 1505 1510 1515

ATT GGC TTT GCT AAG ATG ATA CCA GGA TTC AGA GAC CTC ACC TCT GAG 886 He Gly Phe Ala Lys Met He Pro Gly Phe Arg Asp Leu Thr Ser Glu 1520 1525 1530 1535

GAC CAG ATC GTA CTG CTG AAG TCA AGT GCC ATT GAG GTC ATC ATG TTG 934 Asp Gin He Val Leu Leu Lys Ser Ser Ala He Glu Val He Met Leu 1540 1545 1550

CGC TCC AAT GAG TCC TTC ACC ATG GAC GAC ATG TCC TGG ACC TGT GGC 982 Arg Ser Asn Glu Ser Phe Thr Met Asp Asp Met Ser Trp Thr Cys Gly 1555 1560 1565

AAC CAA GAC TAC AAG TAC CGC GTC AGT GAC GTG ACC AAA GCC GGA CAC 1030 Asn Gin Asp Tyr Lys Tyr Arg Val Ser Asp Val Thr Lys Ala Gly His 1570 1575 1580 AGC CTG GAG CTG ATT GAG CCC CTC ATC AAG TTC CAG GTG GGA CTG AAG 1078 Ser Leu Glu Leu He Glu Pro Leu He Lys Phe Gin Val Gly Leu Lys 1585 1590 1595

AAG CTG AAC TTG CAT GAG GAG GAG CAT GTC CTG CTC ATG GCC ATC TGC 1126 Lys Leu Asn Leu His Glu Glu Glu His Val Leu Leu Met Ala He Cys 1600 1605 1610 1615

ATC GTC TCC CCA GAT CGT CCT GGG GTG CAG GAC GCC GCG CTG ATT GAG 1174 He Val Ser Pro Asp Arg Pro Gly Val Gin Asp Ala Ala Leu He Glu 1620 1625 1630 GCC ATC CAG GAC CGC CTG TCC AAC ACA CTG CAG ACG TAC ATC CGC TGC 1222 Ala He Gin Asp Arg Leu Ser Asn Thr Leu Gin Thr Tyr He Arg Cys 1635 1640 1645 CGC CAC CCG CCC CCG GGC AGC CAC CTG CTC TAT GCC AAG ATG ATC CAG 1270 Arg His Pro Pro Pro Gly Ser His Leu Leu Tyr Ala Lys Met He Gin 1650 1655 1660

AAG CTA GCC GAC CTG CGC AGC CTC AAT GAG GAG CAC TCC AAG CAG TAC 1318 Lys Leu Ala Asp Leu Arg Ser Leu Asn Glu Glu His Ser Lys Gin Tyr 1665 1670 1675

CGC TGC CTC TCC TTC CAG CCT GAG TGC AGC ATG AAG CTA ACG CCC CTT 1366 Arg Cys Leu Ser Phe Gin Pro Glu Cys Ser Met Lys Leu Thr Pro Leu 1680 1685 1690 1695

GTG CTC GAA GTG TTT GGC AAT GAG ATC TCC TGA CTAGGACAGC CTGTGCGGTG 1419 Val Leu Glu Val Phe Gly Asn Glu He Ser * 1700 1705

CCTGGGTGGG GCTGCTCCTC CAGGGCCACG TGCCAGGCCC CGGGCTGGCG GCTACTCAGC 1479

AGCCCTCCTC ACCCGTCTGG GGTTCAGCCC CTCCTCTGCC ACCTCCCCTA TCCACCCAGC 1539 CCATTCTCTC TCCTGTCCAA CCTAACCCCT TTCCTGCGGG CTTTTCCCCG GTCCCTTGAG 1599

ACCTCAGCCA TGAGGAGTTG CTGTTTGTTT GACAAAGAAA CCCAAGTGGG GGCAGAGGGC 1659

AGAGGCTGGA GGCAGGCCTT GCCCAGAGAT GCCTCCACCG CTGCCTAAGT GGCTGCTGAC 1719

TGATGTTGAG GGAACAGACA GGAGAAATGC ATCCATTCCT CAGGGACAGA GACACCTGCA 1779

CCTCCCCCCA CTGCAGGCCC CGCTTGTCCA GCGCCTAGTG GGGTCTCCCT CTCCTGCCTT 1839 ACTCACGATA AATAATCGGC CCACAGCTCC CACCCCACCC CCTTCAGTGC CCACCAACAT 1899

CCCATTGCCC TGGTTATATT CTCACGGGCA GTAGCTGTGG TGAGGTGGGT TTTCTTCCCA 1959

TCACTGGAGC ACCAGGCACG AACCCACCTG CTGAGAGACC CAAGGAGGAA AAACAGACAA 2019

AAACAGCCTC ACAGAAGAAT ATGACAGCTG TCCCTGTCAC CAAGCTCACA GTTCCTCGCC 2079

CTGGGTCTAA GGGGTTGGTT GAGGTGGAAG CCCTCCTTCC ACGGATCCAT GTAGCAGGAC 2139 TGAATTGTCC CCAGTTTGCA GAAAAGCACC TGCCGACCTC GTCCTCCCCC TGCCAGTGCC 2199

TTACCTCCTG CCCAGGAGAG CCAGCCCTCC CTGTCCTCCT CGGATCACCG AGAGTAGCCG 2259

AGAGCCTGCT CCCCCACCCC CTCCCCAGGG GAGAGGGTCT GGAGAAGCAG TGAGCCGCAT 2319

CTTCTCCATC TGGCAGGGTG GGATGGAGGA GAAGAATTTT CAGACCCCAG CGGCTGAGTC 2379

ATGATCTCCC TGCCGCCTCA ATGTGGTTGC AAGGCCGCTG TTCACCACAG GGCTAAGAGC 2439 TAGGCTGCCG CACCCCAGAG TGTGGGAAGG GAGAGCGGGG CAGTCTCGGG TGGCTAGTCA 2499 GAGAGAGTGT TTGGGGGTTC CGTGATGTAG GGTAAGGTGC CTTCTTATTC TCACTCCACC 2559

ACCCAAAAGT CAAAAGGTGC CTGTGAGGCA GGGGCGGAGT GATACAACTT CAAGTGCATG 2619

CTCTCTGCAG GTCGAGCCCA GCCCAGCTGG TGGGAAGCGT CTGTCCGTTT ACTCCAAGGT 2679

GGGTCTTTGT GAGAGTGAGC TGTAGGTGTG CGGGACCGGT ACAGAAAGGC GTTCTTCGAG 2739

GTGGATCACA GAGGCTTCTT CAGATCAATG CTTGAGTTTG GAATCGGCCG CATTCCCTGA 2799

GTCACCAGGA ATGTTAAAGT CAGTGGGAAC GTGACTGCCC CAACTCCTGG AAGCTGTGTC 2859

CTTGCACCTG CATCCGTAGT TCCCTGAAAA CCCAGAGAGG AATCAGACTT CACACTGCAA 2919 GAGCCTTGGT GTCCACCTGG CCCCATGTCT CTCAGAATTC TTCAGGTGGA AAAACATCTG 2979

AAAGCCACGT TCCTTACTGC AGAATAGCAT ATATATCGCT TAATCTTAAA TTTATTAGAT 3039

ATGAGTTGTT TTCAGACTCA GACTCCATTT GTATTATAGT CTAATATACA GGGTAGCAGG 3099

TACCACTGAT TTGGAGATAT TTATGGGGGG AGAACTTACA TTGTGAAACT TCTGTACATT 3159

AATTATTATT GCTGTTGTTA TTTTACAAGG GTCTAGGGAG AGACCCTTGT TTGATTTTAG 3219 CTGCAGAACT GTATTGGTCC AGCTTGCTCT TCAGTGGGAG AAAAACACTT GTAAGTTGCT 3279

AAACGAGTCA ATCCCCTCAT TCAGGAAAAC TGACAGAGGA GGGCGTGACT CACCCAAGCC 3339

ATATATAACT AGCTAGAAGT GGGCCAGGAC AGGCCGGGCG CGGTGGCTCA CGCCTGTAAT 3399

CCCAGCAGTT TGGGAGGTCG AGGTAGGTGG ATCACCTGAG GTCGGGAGTT CGAGACCAAC 3459

CTGACCAACA TGGAGAAACC CTGTCTCTAT TAAAAATACA AAAAAAAAAA AAAAAAAAAA 3519 TAGCCGGGCA TGGTGGCGCA AGCCTGTAAT CCCAGCTACT CAGGAGGCTG AGGCAGAAGA 3579

ATTGAACCCA GGAGGTGGAG GTTGCAGTGA GCTGAGATCG TGCCGTTACT CTCCAACCTG 3639

GACAACAAGA GCGAAACTCC GTCTTAGAAG TGGACCAGGA CAGGACCAGA TTTTGGAGTC 3699

ATGGTCCGGT GTCCTTTTCA CTACACCATG TTTGAGCTCA GACCCCCACT CTCATTCCCC 3759

AGGTGGCTGA CCCAGTCCCT GGGGGAAGCC CTGGATTTCA GAAAGAGCCA AGTCTGGATC 3819 TGGGACCCTT TCCTTCCTTC CCTGGCTTGT AACTCCACCA AGCCCATCAG AAGGAGAAGG 3879

AAGGAGACTC ACCTCTGCCT CAATGTGAAT CAGACCCTAC CCCACCACGA TGTGCCCTGG 3939

CTGCTGGGCT CTCCACCTCA GGCCTTGGAT AATGCTGTTG CCTCATCTAT AACATGCATT 3999

TGTCTTTGTA ATGTCACCAC CTTCCCAGCT CTCCCTCTGG CCCTGCTTCT TCGGGGAACT 4059

CCTGAAATAT CAGTTACTCA GCCCTGGGCC CCACCACCTA GGCCACTCCT CCAAAGGAAG 4119 TCTAGGAGCT GGGAGGAAAA GAAAAGAGGG GAAAATGAGT TTTTATGGGG CTGAACGGGG 4179 AGAAAAGGTC ATCATCGATT CTACTTTAGA ATGAGAGTGT GAAATAGACA TTTGTAAATG 4239

TAAAACTTTT AAGGTATATC ATTATAACTG AAGGAGAAGG TGCCCCAAAA TGCAAGATTT 4299

TCCACAAGAT TCCCAGAGAC AGGAAAATCC TCTGGCTGGC TAACTGGAAG CATGTAGGAG 4359

AATCCAAGCG AGGTCAACAG AGAAGGCAGG AATGTGTGGC AGATTTAGTG AAAGCTAGAG 4419

ATATGGCAGC GAAAGGATGT AAACAGTGCC TGCTGAATGA TTTCCAAAGA GAAAAAAAGT 4479

TTGCCAGAAG TTTGTCAAGT CAACCAATGT AGAAAGCTTT GCTTATGGTA ATAAAAATGG 4539

CTCATACTTA TATAGCACTT ACTTTGTTTG CAAGTACTGC TGTAAATAAA TGCTTTATGC 4599 AAACC 4604

(2) INFORMATION FOR SEQ ID NO: 16: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 428 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 16 :

Met Glu Ala Met Ala Ala Ser Thr Ser Leu Pro Asp Pro Gly Asp Phe 1 5 10 15

Asp Arg Asn Val Pro Arg He Cys Gly Val Cys Gly Asp Arg Ala Thr 20 25 30 Gly Phe His Phe Asn Ala Met Thr Cys Glu Gly Cys Lys Gly Phe Phe 35 40 45

Arg Arg Ser Met Lys Arg Lys Ala Leu Phe Thr Cys Pro Phe Asn Gly 50 55 60

Asp Cys Arg He Thr Lys Asp Asn Arg Arg His Cys Gin Ala Cys Arg 65 70 75 80

Leu Lys Arg Cys Val Asp He Gly Met Met Lys Glu Phe He Leu Thr 85 90 95

Asp Glu Glu Val Gin Arg Lys Arg Glu Met He Leu Lys Arg Lys Glu 100 105 110 Glu Glu Ala Leu Lys Asp Ser Leu Arg Pro Lys Leu Ser Glu Glu Gin 115 120 125

Gin Arg He He Ala He Leu Leu Asp Ala His His Lys Thr Tyr Asp 130 135 140 Pro Thr Tyr Ser Asp Phe Cys Gin Phe Arg Pro Pro Val Arg Val Asn 145 150 155 160

Asp Gly Gly Gly Ser His Pro Ser Arg Pro Asn Ser Arg His Thr Pro 165 170 175

Ser Phe Ser Gly Asp Ser Ser Ser Ser Cys Ser Asp His Cys He Thr

180 185 190 Ser Ser Asp Met Met Asp Ser Ser Ser Phe Ser Asn Leu Asp Leu Ser 195 200 205

Glu Glu Asp Ser Asp Asp Pro Ser Val Thr Leu Glu Leu Ser Gin Leu 210 215 220

Ser Met Leu Pro His Leu Ala Asp Leu Val Ser Tyr Ser He Gin Lys 225 230 235 240

Val He Gly Phe Ala Lys Met He Pro Gly Phe Arg Asp Leu Thr Ser 245 250 255

Glu Asp Gin He Val Leu Leu Lys Ser Ser Ala He Glu Val He Met 260 265 270 Leu Arg Ser Asn Glu Ser Phe Thr Met Asp Asp Met Ser Trp Thr Cys

275 280 285

Gly Asn Gin Asp Tyr Lys Tyr Arg Val Ser Asp Val Thr Lys Ala Gly 290 295 300

His Ser Leu Glu Leu He Glu Pro Leu He Lys Phe Gin Val Gly Leu 305 310 315 320

Lys Lys Leu Asn Leu His Glu Glu Glu His Val Leu Leu Met Ala He 325 330 335

Cys He Val Ser Pro Asp Arg Pro Gly Val Gin Asp Ala Ala Leu He 340 345 350 Glu Ala He Gin Asp Arg Leu Ser Asn Thr Leu Gin Thr Tyr He Arg 355 360 365

Cys Arg His Pro Pro Pro Gly Ser His Leu Leu Tyr Ala Lys Met He 370 375 380

Gin Lys Leu Ala Asp Leu Arg Ser Leu Asn Glu Glu His Ser Lys Gin 385 390 395 400

Tyr Arg Cys Leu Ser Phe Gin Pro Glu Cys Ser Met Lys Leu Thr Pro 405 410 415

Leu Val Leu Glu Val Phe Gly Asn Glu He Ser * 420 425

Claims

What is claimed is:

1. A method comprising, identifying individuals having a certain phenotype, determining the presence or absence of gene markers associated with the phenotype, and instituting a lifestyle change to exploit or counteract the phenotype expressed by the gene marker.

2. The method of claim 1 wherein the phenotype is low level HDL-C and HDI^-C, the gene marker is APO E2 and exercise training is instituted to improve the HDL-C and

HDL₂-C levels.

3. The method of claim 1 wherein the phenotype is hypertension, the gene marker is at least one insertion ("I") ACE allele and exercise training is instituted to decrease systolic and diastolic blood pressure.

4. The method of claim 1 wherein the phenotype is a reduction of bone mineral density, the gene marker is BB vitamin D receptor and exercise training is instituted to increase bone mineral density.

5. The method of claim 1 wherein the phenotype is a determined VQ,max value for a determined habitual level of physical activity, the gene marker is at the angiotensinconverting enzyme locus and a specific aerobic sport is conducted to exploit, for sport, the presence of the marker.

6. A method of determining the risk of cardiovascular disease in an individual comprising, determining the ACE phenotype of the individual.

7. The method of claim 1 wherein the phenotype is hypertension, the gene marker is located at restriction cites of the lipoprotein lipase gene locus and exercise training is instituted to decrease blood pressure.

8. The method of claim 7 wherein the gene marker is at the LPL PvuII restriction site.

9. The method of claim 7 wherein the gene marker is at the Hindlll restriction site.